Indolopyrrolocarbazole derivatives

ABSTRACT

Indolopyrrocarbazole derivatives represented by formula (I) and the pharmaceutically acceptable salts thereof have excellent antitumor activity as evidenced by their in vitro proliferation inhibiting activity against mouse leukemia cell, human gastric cancer cell, human lung cancer cell and human colon cancer cell, ##STR1## wherein R 1  and R 2  independently represent, for example, a hydrogen atom or various hydrocarbon groups which may be substituted or heterocyclic groups which may also be substituted; or a group --Y--R 3  where Y represents a carbonyl group, thiocarbonyl group or sulfonyl group and R 3  represents a hydrogen atom or one of various aliphatic, cycloaliphatic, aryl, nitrogen-containing (e.g. amino, hydrazino, etc) or heterocyclic groups, which groups may be substituted by various substituents; or R 1  and R 2  may combine to represent a lower alkylidene group which may be substituted; or R 1  and R 2 , together with the N-atom to which they are bonded form a heterocyclic group which may be substituted; 
     G represents a pentose or hexose group; and X 1  and X 2 , independently, represent, for example, hydrogen, halogen, amino, hydroxyl, alkoxy, aryloxy, carboxyl, alkoxycarbonyl or alkyl. These compounds have improved water solubility as compared to rebeccamycin.

This is a division of application Ser. No. 08/255,980, filed Jun. 8,1994, now U.S. Pat. No. 5,591,842; which is a continuation-in-part ofapplication Ser. No. 07/981,070 filed Nov. 24, 1992.

This invention is useful in the field of medicine, and relates to novelindolopyrrolocarbazole derivatives inhibiting proliferation of antitumorcells and exhibiting an antitumor effect, a process for preparationthereof and a use thereof.

In the field of cancer chemotherapy, many compounds are already put topractical use as an antitumor agent. However, the effect thereof onvarious kind of tumors is not always adequate, and the problem ofresistance of tumor cells against these drugs makes clinical use ofthese antitumor agents complicated [refer to The 47th Japan Society ofCancer General Meeting Article, pages 12 to 15 (1988)].

In such state of things, development of novel carcinostatic substancesis always made in the filed of cancer therapy. Particularly, substancesare necessitated which overcome resistance against existingcarcinostatic substances and exhibit effectiveness against such kinds ofcancers on which existing carcinostatic substances cannot exhibitsufficient effects.

In the light of such present state of things, the present inventorswidely screened microbial metabolic products, as a result, found a novelantitumor activity-possessing compound BE-13793C(12,13-dihydro-1,11-dihydroxy-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione),and disclosed it [refer to Japanese Laid-Open Patent Publication No.20277/1991 and J. Antibiotics, 44, 723-728 (1991)].

Thereafter, the present inventors created indolopyrrolocarbazolecompounds having an excellent antitumor activity by chemically modifyingBE-13793C, and disclosed them (refer to PCT/WO91/18003).

For the purpose of creating compounds having a further excellentantitumor activity by chemically modifying previously disclosedindolopyrrolocarbazole antitumor compounds, the present inventorssynthesized many indolopyrrolocarbazole derivatives, investigated theirantitumor activity, and as a result, now, found that a series ofcompounds represented by the following general formula are novelcompounds having an extremely excellent antitumor activity.

Thus, this invention provides indolopyrrolocarbazole derivativesrepresented by the following general formula and pharmaceuticallyacceptable salts thereof. ##STR2## wherein

R¹ and R² each independently represent a hydrogen atom, lower alkylgroup, lower alkenyl group, lower alkynyl group, aryl group, aralkylgroup or heterocyclic group (the lower alkyl group, lower alkenyl group,lower alkynyl group, aryl group, aralkyl group and heterocyclic groupmay each have 1 to 5 substituents selected from the group consisting ofcarboxyl groups, carbamoyl groups, sulfo groups, amino groups, cyanogroups, mono-lower alkylamino groups, di-lower alkylamino groups,hydroxyl groups and halogen atoms), or a group of the formula --Y--R³,and therein Y represents a carbonyl group, thiocarbonyl group orsulfonyl group, and R³ represents a hydrogen atom, lower alkyl group,cycloalkyl group, cycloalkylalkyl group, aryl group, aralkyl group,lower alkoxy group, hydrazino group, amino group, arylamino group,carbamoyl group or heterocyclic group (the lower alkyl group, cycloalkylgroup, cycloalkylalkyl group, aryl group, aralkyl group and heterocyclicgroup may each have 1 to 4 substituents selected from the groupconsisting of halogen atoms, optionally protected hydroxyl groups, aminogroups, carboxyl groups, carbamoyl groups, cyano groups and loweralkoxycarbonyl groups, and the amino group and carbamoyl group may eachbe mono- or di-substituted by lower alkyl group(s) optionallysubstituted by substituent(s) selected from the group consisting ofhalogen atoms, hydroxyl groups, amino groups, carboxyl groups, carbamoylgroups and lower alkoxycarbonyl groups); or

R¹ and R² combine to represent a lower alkylidene group (the loweralkylidene group may have 1 to 4 substituents selected from the groupconsisting of amino groups, mono-lower alkylamino groups, di-loweralkylamino groups, hydroxyl groups, carboxyl groups and sulfonylgroups); or

R¹ and R² combine together with the nitrogen atom to which they bind toform a heterocyclic group (the heterocyclic group may have on the ringlower alkyl group(s) optionally substituted by group(s) selected fromthe group consisting of amino groups, hydroxyl groups, a carboxyl groupsand sulfo groups),

G represents a pentose group or hexose group, and

X¹ and X² each independently represent a hydrogen atom, halogen atom,amino group, mono-lower alkylamino group, di-lower alkylamino group,hydroxyl group, lower alkoxy group, aralkoxy group, carboxyl group,lower alkoxycarbonyl group or lower alkyl group.

The term of "lower" used in the present invention means that the carbonnumber of the group or compound to which this term is attached is 6 orless, preferably 4 or less.

The "lower alkyl group" is a straight-chain or branched chain alkylgroup having 1 to 6 carbon atoms, and examples thereof are a methylgroup, an ethyl group, a propyl group, an isopropyl group, a butylgroup, an isobutyl group, a sec-butyl group, a tert-butyl group, apentyl group, an isopentyl group, a neopentyl group, a hexyl group, etc.

The "lower alkenyl group" includes a straight-chain or branched chainalkenyl group having 3 to 6 carbon atoms, and examples thereof are apropenyl group, a 2-butenyl group, a 3-butenyl group, a 3-pentenylgroup, a 4-hexenyl group, etc.

The "lower alkynyl group" can be a straight-chain or branched chainalkynyl group having 3 to 6 carbon atoms, and examples thereof are apropynyl group, a 2-butynyl group, a 3-butynyl group, a 3-pentynylgroup, a 4-hexynyl group, etc.

The "cycloalkyl group" includes a 3- to 6-membered cycloalkyl group, andexamples thereof are a cyclopropyl group, a cyclobutyl group, acyclopentyl group, a cyclohexyl group, etc.

The "cycloalkyl lower alkyl group" means an alkyl group substituted by acycloalkyl group wherein the cycloalkyl and lower alkyl parts have theabove meanings, respectively, and examples thereof are acyclopropylmethyl group, a cyclobutylmethyl group, a cyclopentylmethylgroup, a cyclohexylmethyl group, a 1-cyclopropylethyl group, a2-cyclopropylethyl group, a 1-cyclobutylethyl group, a 2-cyclobutylethylgroup, a 1-cyclopentylethyl group, a 2-cyclopentylethyl group, a1-cyclohexylethyl group, a 3-cyclohexylpropyl group, a3-cyclopentylpropyl group, a 4-cyclohexylbutyl group, a4-cyclopentylbutyl group, etc., and preferably, the cycloalkylalkylgroup has 4 to 10 carbon atoms in total.

The "aryl group" can be monocyclic or polycyclic, and aryl groups having6 to 12 carbon atoms can be mentioned such as a phenyl group, a naphthylgroup and a tetrahydronaphthyl group.

The "aralkyl" group means a lower alkyl group substituted by an arylgroup wherein the aryl and lower alkyl parts have the above meanings,respectively, and aralkyl groups having 7 to 15 carbon atoms can bementioned such as, for example, a benzyl group, a phenethyl group, aphenylpropyl group, a phenylbutyl group, a phenylpentyl group, anaphthylmethyl group and a naphthylethyl group.

The "heterocyclic group" includes a 5- or 6-membered heterocyclic groupcontaining 1 to 4 hetero atoms selected from the group consisting ofnitrogen atoms, oxygen atoms and sulfur atoms, and there can bementioned aromatic heterocyclic groups such as, for example, a pyrrolylgroup, a furyl group, a thienyl group, an oxazolyl group, an isoxazolylgroup, a thiazolyl group, an isothiazolyl group, an imidazolyl group, apyrazolyl group, an oxadiazolyl group, a thiadiazolyl group, a triazolylgroup, a tetrazolyl group, a furazanyl group, a pyridyl group, apyridazinyl group, a pyrimidinyl group, a pyrazinyl group and atriazinyl group; and nonaromatic heterocyclic groups such as, forexample, a dihydrothienyl group, a tetrahydrothienyl group, a pyrrolinylgroup, a pyrrolidinyl group, an imidazolidinyl group, an imidazolinylgroup, a piperidinol group, a piperazinyl group, an oxazolinyl group, anoxazolidinyl group, an isoxazolinyl group, an isoxazolidinyl group, athiazolinyl group, a thiazolidinyl group, an isothiazolinyl group, anisothiazolidinyl group, a 1,2-dithiolanyl group, a 1,3-dithiolanylgroup, a 1,2-dithiolyl, a 1,3-dithiolyl group, a dihydrothiopyranylgroup, a tetrahydrothiopyranyl group, a 1,4-dithianyl group, a1,4-dithiinyl group, a 1,4-oxathiinyl group and a thiomorpholinyl group.

As the "mono-lower alkylamino groups", there can, for example, bementioned a methylamino group, an ethylamino group, a propylamino group,an isopropylamino group, a butylamino group, a pentylamino group, ahexylamino group, etc., and as the "di-lower alkylamino groups" therecan, for example, be mentioned a dimethylamino group, anethylmethylamino group, a diethylamino group, an ethylpropylamino group,a dipropylamino group, a butylmethylamino group, a dibutylamino group, abutylethylamino group, a methylpentylamino group, a hexylmethylaminogroup, an ethylhexylamino group, etc.

The "arylamino group" means an amino group substituted by an aryl groupwherein the aryl part has the above meanings, and the arylamino groupcan be mentioned such as, for example, a phenylamino group and anaphthylamino group.

The "halogen atoms" include a fluorine atom, a chlorine atom, a bromineatom and an iodine atom.

As the "lower alkylidene groups", there can be mentioned straight-chainor branched alkylidene groups having 1 to 6 carbon atoms such as, forexample, a methylene group, an ethylidene group, a propylidene group, anisopropylidene group, a butylidene group, an isobutylidene group, asec-butylidene group, a pentylidene group, an isopentylidene group, aneopentylidene group and a hexylidene group.

The "lower alkoxy group" means a (lower alkyl)-O-group wherein the loweralkyl part has the above meaning, and examples thereof are a methoxygroup, an ethoxy group, a propoxy group, an isopropoxy group, a butoxygroup, an isobutoxy group, a sec-butoxy group, a tert-butoxy group, apentoxy group, an isopentoxy group, a neopentoxy group, a hexoxy group,etc.

The "lower alkoxycarbonyl group" means a (lower alkoxy)-CO-group whereinthe lower alkoxy part has the above meaning, and examples thereof are amethoxycarbonyl group, an ethoxycarbonyl group, a propyloxycarbonylgroup, an isopropyloxycarbonyl group, a butyloxycarbonyl group, anisobutyloxycarbonyl group, a pentyloxycarbonyl group, a hexyloxycarbonylgroup, etc.

The "aralkoxy group" means a lower alkoxy group substituted by an arylgroup wherein the aryl and lower alkoxy parts have the aforesaidmeanings, respectively, and examples thereof are a benzyloxy group, aphenethyloxy group, a phenylpropoxy group, an α-naphthylmethoxy group, aβ-naphthylmethoxy group, a naphthylethoxy group, atetrahydronaphthylmethoxy group, etc.

Mentioned as examples of the protective group in the "optionallysubstituted hydroxyl group" are alkanoyl groups having 2 to 6 carbonatoms such as an acetyl group, a propronyl group and a butyryl group;aroyl groups such as a benzoyl group; substituted or unsubstitutedaralkyl groups such as a benzyl group and a 4-methoxybenzyl group;groups forming an acetal such as acetonide; etc.

The "pentose group" and "hexose group" mean a pentose group and a hexosegroup the hydroxyl groups of which may be substituted by the same ordifferent 1 to 3 groups selected from the group consisting of hydrogenatoms, lower alkyl groups, lower alkylcarbonyloxy groups, lower alkoxygroups and amino groups, or oxidized, and there can be mentioned groupsderived from pentoses such as, for example, ribose, arabinose, xyloseand 2-deoxyribose, and groups derived from hexoses such as, for example,allose, glucose, mannose, galactose, glucosamine, galactosamine,2-deoxyglucose, 4-O-methylglucose, rhamnose and glucuronic acid.

Preferred among the compounds of the aforesaid formula [I] provided bythis invention are compounds represented by the following formula##STR3## wherein

R¹¹ and R²¹ each independently represent a hydrogen atom, lower alkylgroup, lower alkenyl group, aryl group, aralkyl group, pyrrolyl group,oxazolyl group, isoxazolyl group, thiazolyl group, imidazolyl group,pyridyl group, pyrimidinyl group, oxazolinyl group, oxazolidinyl group,imidazolinyl group, imidazolidinyl group, pyrrolidinyl group,piperazinyl group, thiazinyl group, thiazolidinyl group (the lower alkylgroup, lower alkenyl group, aryl group, aralkyl group and heterocyclicgroup may have 1 to 5 substituents selected from the group consisting ofcarboxyl groups, carbamoyl groups, cyano groups and hydroxyl groups), ora group of the formula --Y--R³¹, and therein Y represents a carbonylgroup, thiocarbonyl group or sulfonyl group, and R³¹ represents ahydrogen atom, lower alkyl group, aryl group (the lower alkyl group andaryl group may have 1 to 4 substituents selected from the groupconsisting of halogen atoms, optionally protected hydroxyl groups, aminogroups and carboxyl groups), amino group, hydrazino group, arylaminogroup, lower alkoxy group, carbamoyl group, pyrrolyl group, oxazolylgroup, isoxazolyl group, thiazolyl group, imidazolyl group, pyridylgroup, pyrimidinyl group, oxazolinyl group, oxazolidinyl group,imidazolinyl group, imidazolidinyl group, pyrrolidinyl group,piperazinyl group, thiazinyl group or thiazolidinyl group; or

R¹¹ and R²¹ combine to represent a lower alkylidene group optionallyhaving carboxyl group(s), or

R¹¹ and R²¹ combine together with the nitrogen atom to which they bindto form a pyrrolidinyl group, imidazolidinyl group, imidazolinyl group,piperidino group or, piperazinyl group (these heterocyclic groups mayhave on the ring lower alkyl group(s) optionally substituted by hydroxygroup(s)),

G¹ represents a group of the formula ##STR4## and therein R⁷ representsa hydrogen atom or lower alkyl group and R⁸ represents a hydroxyl groupor amino group, and

X¹¹ and X²¹ bind to the indolopyrrolocarbazole rings at the 1- or2-position and at the 10- or 11-position, respectively, and eachindependently represent a halogen atom, hydroxyl group, lower alkoxygroup or aralkoxy group.

Further preferred compounds are those represented by the followingformula ##STR5## wherein

R¹² represents a hydrogen atom or lower alkyl group,

R²² represents a hydrogen atom, lower alkyl group (the lower alkyl groupmay have 1 to 5 substituents selected from the group consisting ofcarboxyl groups, carbamoyl groups, hydroxyl groups and cyano groups),aryl group, aralkyl group (the aryl group and aralkyl group may have 1to 4 substituents selected from the group consisting of hydroxyl groupsand carboxyl groups), pyridyl group, imidazolyl group, imidazolinylgroup, thiazolyl group, pyrrolidinyl group, piperazinyl group, or agroup of the formula --Y--R³², and therein Y represents a carbonylgroup, thiocarbonyl group or sulfonyl group, and when Y is a carbonylgroup or thiocarbonyl group, R³² represents a hydrogen atom, lower alkylgroup, aryl group (the lower alkyl group and aryl group may have 1 to 4substituents selected from the group consisting of halogen atoms,optionally protected hydroxyl groups, amino groups and carboxyl groups),amino group, hydrazino group, arylamino group, lower alkoxy group,carbamoyl group, pyridyl group, pyrimidinyl group, imidazolinyl group orpyrrolidinyl group, and when Y is a sulfonyl group, R³² represents alower alkyl group or aryl group; or

R¹² and R²² combine to represent a lower alkylidene group havingcarboxyl group(s); or

R¹² and R²² combine together with the nitrogen atom to which they bindto form a pyrrolidinyl group, piperidino group or piperazinyl group(these heterocyclic groups may have on the ring lower alkyl group(s)optionally having hydroxyl group(s)), and

G¹, X¹¹ and X²¹ have the same meanings as defined in the above formula[Ia].

Preferred as G¹ is generally ##STR6## and preferred as X¹¹ and X²¹ arehydroxyl groups bound to the 1-position and 11-position of theindolopyrrolocarbazole ring, respectively.

An especially preferred class of compounds of formula [Ib] are those inwhich R²² represents the lower alkyl (C₁ -C₆) group substituted with upto five hydroxyl groups. Further preferred are those wherein X¹¹ and X²¹each represent a hydroxyl group, especially those in which the hydroxylgroups are positioned at the 1- and 11-positions or at the 2- and10-positions of the indolopyrrolocarbazole rings.

An especially preferred class of compounds within the scope of formula[I] and formula [Ib] are those represented by the following formula##STR7## wherein R represents a branched or straight chain alkyl groupof 2 to 5 carbon atoms containing 1 to 4 hydroxyl groups.

Preferred as R are branched or straight-chain alkyl groups of 2 or 3carbon atoms containing 1 or 2 hydroxyl groups, such as, for example,--CH₂ CH₂ OH, --CH₂ CH(OH)CH₂ OH, --CH(CH₂ OH)₂ and the like.

The compounds of this Invention can exist in the form of pharmaceuticalacceptable salts. Such salts include addition salts with inorganic acidssuch as, for example, hydrochloric acid and sulfuric acid, and withorganic acids such as, for example, acetic acid, citric acid, tartaricacid and maleic acid. Further, in case the compounds of this inventioncontain an acidic group, the acidic group can exist in the form ofalkali metal salts such as, for example, a potassium salt and a sodiumsalt; alkaline earth metal salts such as, for example, a magnesium saltand a calcium salt; and salts with organic bases such as an ethylaminesalt and an arginine salt.

A compound of the formula [I] set forth in this invention can beprepared by reacting a compound represented by the following formula ora derivative thereof wherein the functional groups are protected##STR8## or ##STR9## wherein,

Z represents a hydrogen atom or substituted or unsubstituted lower alkylgroup, and X¹, X² and G have the same meanings as defined above

with a compound represented by the following general formula or aderivative thereof wherein in case R¹³ and R²³ contain a functionalgroup, the functional group is each protected ##STR10## wherein

R¹³ and R²³ each independently represent a hydrogen atom, lower alkylgroup, lower alkenyl group, lower alkynyl group, aryl group, aralkylgroup or heterocyclic group (the lower alkyl group, lower alkenyl group,lower alkynyl group, aryl group, aralkyl group and heterocyclic groupmay have 1 to 5 substituents selected from the group consisting ofcarboxyl groups, carbamoyl groups, sulfo groups, amino groups, cyanogroups, mono-lower alkylamino groups, di-lower alkylamino groups,hydroxyl groups and halogen atoms), or a group of the formula --Y--R³,and herein Y represents a carbonyl group, thiocarbonyl group or sulfonylgroup, and R³ represents a hydrogen atom, lower alkyl group, cycloalkylgroup, cycloalkylalkyl group, aryl group, aralkyl group, lower alkoxygroup, hydrazino group, amino group, arylamino group, or carbamoyl groupor heterocyclic group (the lower alkyl group, cycloalkyl group,cycloalkylalkyl group, aryl group, aralkyl group and heterocyclic groupmay each have 1 to 4 substituents selected from the group consisting ofhalogen atoms, optionally protected hydroxyl groups, amino groups,carboxyl groups, carbamoyl groups, cyano groups and lower alkoxycarbonylgroups, and the amino group and carbamoyl group may each be mono- ordi-substituted by lower alkyl group(s) optionally substituted bygroup(s) selected from the group consisting of halogen atoms, hydroxylgroups, amino groups, carboxyl groups, carbamoyl groups and loweralkoxycarbonyl groups); or

R¹³ and R²³ combine together with the nitrogen atom to which they bindto form a heterocyclic group (the heterocyclic group may have on thering lower alkyl group(s) optionally substituted by group(s) selectedfrom the group consisting of amino groups, hydroxyl groups, carboxylgroups and sulfo groups); if necessary, removing the protective group(s)existing in the product to prepare a compound represented by the generalformula ##STR11##

wherein R¹³, R²³, X¹, X², G have the same meanings as defined above;

or either formylating, alkylating, alkenylating, alkynylating,aralkylating, carbamoylating, thiocarbamoylating, alkanoylating orsulfonylating the amino group ##STR12## of the compound of the aboveformula [Ic] or the derivative thereof wherein the functional groups areprotected when R¹³ and R²³ represent a hydrogen atom, or condensing theabove compound or derivative with a compound represented by thefollowing formula or a derivative thereof wherein a functional group isprotected

    OHC--R.sup.6                                               [V]

wherein R⁶ represents a hydrogen atom or carboxyl group, or a loweralkyl group optionally having 1 to 4 substituents selected from thegroup consisting of amino groups, mono- lower alkylamino groups,di-lower alkylamino groups, hydroxyl groups, carboxyl groups and sulfogroups,

and if necessary, removing the protective groups existing in theproduct; or reducing the double bonds of the compound of the aboveformula [Ic] when R¹³ and/or R²³ contain a double bond or the compoundprepared by condensing the compound [Ic] and the compound [V], or thederivative thereof wherein the functional groups are protected, and ifnecessary removing the protective groups existing in the product; and ifnecessary, converting the resulting compound of the formula [I] into apharmaceutically acceptable salt.

Herein, the terms of alkylation, alkenylation, alkynylation,aralkylation, alkanoylation and sulfonylation are widely interpreted,and mean all of the reactions to introduce substituents corresponding toR¹ and R² in the structure of the compounds of this invention, and forexample, alkylation means introduction of a substituted or unsubstitutedalkyl group included in this invention.

Reaction of a compound of the formula [II] or [III] (hereafter,including a derivative thereof wherein a functional group is introduced)with a compound of the formula [IV] (hereafter, including a derivativethereof wherein its functional groups are protected) can be carried outin accordance with reaction known per se of an imide or acid anhydridewith a hydrazine or a hydrazine derivative, and can, for example, becarried out in the absence of a solvent or in an inert solvent, forexample a solvent such as N,N-dimethylformamide at a temperature betweenabout 0° C. and the boiling point of the solvent, preferably in therange of about room temperature to about 80° C.

The use quantity of the compound of the formula [IV] to the compound ofthe formula [II] or [III] is not particularly limited, and can be variedover a wide range according to the kind of the compound, reactionconditions, etc., but usually, it is suitable to use the compound of theformula [IV] in a quantity in the range of at least 1 mole, preferably 1to 10 moles, particularly 3 to 5 moles per mole of the compound of theformula [II] or [III]. Further, when the compound of the formula [IV] isliquid in the reaction temperature, it is also possible to use thecompound in a largely excessive quantity, for example in a rate of 10 to40 moles per mole of the compound of the formula [II] or [III] so as tomake it serve as a solvent.

Thereby, there can be obtained a compound of the above formula [Ic]wherein the existing functional groups are sometimes protectedappropriately.

The thus obtained compound of the formula [Ic] in case R¹³ and R²³represent a hydrogen atom or a derivative thereof wherein its functionalgroups are protected (hereafter, generally referred to as a compound of[Ic-1]) can be formylated, alkylated, alkenylated, alkynylated,aralkylated, carbamoylated, thiocarbamoylated, alkanoylated orsulfonylated to give a corresponding compound of the formula [Ic] incase at least one of R¹³ and R²³ represents a group except for ahydrogen atom defined on these groups.

Formylation of a compound of the formula [Ic-1] can be carried outaccording to a method usually used in formylation of an amino group, andcan, for example, be carried out by heating it together with formicacid, formamide, dimethylformamide or the like, or by a method to reactit with a mixture of formic acid and an acid anhydride in a solventhaving no bad influence or without any solvent, or by another means.

Reaction of the compound of the formula [Ic-1] with formic acid,formamide, dimethylformamide or the like is usually carried out at atemperature in the range of 30° C. to the boiling point of the solvent,but if necessary, can also be carried out at a temperature above orunder such temperature, and reaction time is usually in the range of 30minutes to 2 days. Preferably, the reaction is carried out usually inthe presence of an acid catalyst such as hydrochloric acid or sulfuricacid.

Formylation using a mixture of formic acid with an acid anhydride isusually carried out at a comparatively low temperature in the range of-5° C. to room temperature, but can, if necessary, be carried out in arange above or under this. Further, reaction time is usually 10 minutesto 5 hours, but can, if necessary, be lengthened or shortened.

Alkylation, alkenylation, alkynylation and aralkylation of a compound ofthe formula [Ic-1] can be carried out in accordance with a method knownper se, for example, reaction with an alkylating agent, alkenylatingagent, alkynylating agent or aralkylating agent such as an alkyl halide,an alkenyl halide, an alkynyl halide, an aralkyl halide, an alkylmesylate, an alkenyl mesylate, an aralkyl mesylate, an alkyl tosylate oran aralkyl tosylate; or a method to condense it with an aldehydecompound or a ketone compound and reduce the resultant condensate; orthe like. The reduction reaction at that time can be carried outaccording to a method using formic acid, a metal or a metal hydride or ausual method such as a catalytic reduction method using palladium-carbonor the like.

Carbamoylation and thiocarbamoylation of a compound of the formula[Ic-1] can be carried out by reacting it with a correspond isocyanatecompound or thioisocyanate compound in the absence of solvent or in asuitable solvent. Reaction temperature can be in the range of about -20°C. to the boiling point of the solvent, preferably about 0° to about 50°C.

Alkanoylation of a compound of the formula [Ic-1] can be carried out bya method to react it with a corresponding acid halide or an acidanhydride in the absence of a solvent or in a suitable solvent. Reactioncan usually be carried out at a temperature in the range of about -5° C.to the boiling point of the solvent, and if necessary, can also becarried out at a temperature below this.

The acid halide or acid anhydride is, usually, used in a rate of smallexcess to the compound of the formula [Ic-1], but can, if necessary, beused in a quantity below or above this, and reaction time can, usually,be 30 minutes to 2 days.

Sulfonylation of a compound of the formula [Ic-1] can be carried out byreacting it with a reagent such as a corresponding organic sulfonic acidanhydride or organic sulfonyl halide in the presence or absence of abase. Reaction temperature can, usually, be sufficient in the range ofabout -10° C. to about 50° C., but can, if necessary, be a temperatureabove or under this, and reaction time can, usually, be 30 minutes to 3days. A reagent such as an organic sulfonic acid anhydride or an organicsulfonyl halide is, usually, used in a rate of small excess, but canalso be used, in a quantity above or under this.

Further, condensation reaction of a compound of the formula [Ic-1] witha compound of the above formula [V] (including a derivative thereofwherein the functional groups are protected) is so-called Schiff baseformation reaction, and can, for example, usually be carried out in asolvent inert to the reaction, e.g. in a solvent such astetrahydrofuran, at a temperature between about 0° C. to the boilingpoint of the solvent, preferably in the range of room temperature toabout 50° C. Reaction time is usually in the range of 30 minutes to 2days, but can, if necessary, be a time above or under this.

Use quantity of the compound of the formula [V] to the compound of theformula [Ic-1] is not strictly limited, but usually, it is suitable touse the compound of the formula [V] in a rate of 1 to 50 moles,particularly 3 to 10 moles per mole of the compound of the formula[Ic-1].

The hydrazone compound obtained by the above reaction can be subjectedto usual catalytic hydrogenation reaction using palladium-carbon or thelike to give a compound of the formula [I] wherein R¹ or R² represents ahydrogen atom.

In the foregoing processes, protection of functional groups in rawmaterial compounds and removal of protective groups existing in theformed compounds can be carried out using usual and optional methodswidely known in the chemical field.

Further, isolation and purification of compounds produced by the abovereactions can be carried out according to methods known per se in thefield of organic synthetic chemistry, for example precipitation methods,solvent extraction methods, recrystallization, chromatography, etc.

A compound of the above formula [II] used as a starting raw material inthe aforesaid processes can be prepared by glycosidating a compoundrepresented by the general formula ##STR13##

wherein X¹, X² and Z have the same meanings as defined above

prepared by a process known per se [refer to J. Chem. Soc. PerkinTransactions I. pp 2475-2480 (1990)] or a derivative thereof wherein thefunctional groups are protected.

Glycosidation of the compound of the formula [VI] or the derivativethereof wherein the functional groups are protected can be carried outby a process known per se [refer to J. Am. Chem. Soc. 60, 2559 (1938)],for example by condensing it with a reactive derivative of a pentose orhexose wherein the hydroxyl groups are protected, e.g.1-bromo-2,3,4,6-O-tetraacetylglucose, using as an activating agentmercury cyanide, silver carbonate, silver oxide or the like, preferablysilver oxide, in an aprotic solvent, e.g. a solvent such as benzene,toluene or methylene chloride at a temperature of about 0° C. to about100° C., preferably about 80° C.

Alternatively, a compound of the formula [II] can also be preparedaccording to the process disclosed in the aforesaid PCT/WO91/18003.

Further, a compound of the formula [III] can be prepared by treatingwith a base a thus obtained compound of the formula [II] or derivativethereof wherein the functional groups are protected.

Preferred as the base is an aqueous solution of potassium hydroxide, andtreatment with this base can usually be carried out at room temperature,but in some cases can also be carried out with heating up to atemperature of about 50° C.

Neutralization or acidification of the reaction mixture can, ifnecessary, be carried out using hydrochloric acid, and thereby it ispossible to precipitate the compound of the formula [III] as crystals.

The compounds of formula [II] in which G represents a glycosyl group offormula ##STR14## (these glycosyl group containing compounds may bereferred to as compounds of formula [VII]) may, preferably, be preparedby a fermentation culturing reaction of a compound of the generalformula ##STR15## as described in detail in commonly assigned priorcopending application Ser. No. 08/068,097 filed May 28, 1993, on pages20-25 and in Examples C and D, the entire disclosure of which isincorporated herein in its entirety by reference thereto.

The compounds of the formula [I] provided by this invention have anexcellent antitumor action as shown in the following pharmacologicaltest examples.

(1) Therapeutic effect against mouse tumor (P388)

Therapeutic effect of the compounds of this invention against mousetumor (P388) is shown in Tables 1 and 2.

                  TABLE I                                                         ______________________________________                                        Effect of the compound of Example 2 against P388                                       Dose.sup.(2), i.p                                                    Tumor.sup.(1)                                                                          (mg/kg/injection)                                                                           MST.sup.(3) (day)                                                                        T/C (%).sup.(4)                             ______________________________________                                        P388     0             12.3 ± 1.06                                                                           100                                                  1             15.8 ± 0.84                                                                           128                                                  3             17.8 ± 1.92                                                                           145                                                  10            >26.4 ± 18.82                                                                         >245                                                 30            >42.2 ± 24.38                                                                         >343                                                 100           >47.2 ± 18.90                                                                         >384                                        ______________________________________                                    

                  TABLE 2                                                         ______________________________________                                        Effect of the compound of Example 5 against P388                                       Dose.sup.(2), i.p                                                    Tumor.sup.(1)                                                                          (mg/kg/injection)                                                                           MST.sup.(3) (day)                                                                        T/C (%).sup.(4)                             ______________________________________                                        P388     0             12.3 ± 0.95                                                                           100                                                  1             16.8 ± 0.84                                                                           137                                                  3             17.8 ± 1.92                                                                           145                                                  10            >26.2 ± 10.18                                                                         >213                                                 30            >23.4 ± 9.74                                                                          >190                                                 100           >36.4 ± 8.05                                                                          >296                                        ______________________________________                                         (Footnotes of Tables 1 and 2)                                                 .sup.(1) Tumor inoculation: 10.sup.6  cancer cells were intraperitoneally     inoculated.                                                                   .sup.(2) Dose: After tumor inoculation, each dose was intraperitoneally       administered once a day from the 1st day to 10th day.                         .sup.(3) MST: mean survival number of days                                    .sup.(4) T/C (%): (MST of treatment group/MST of control) × 100         .sup.(5) Standard: in case of T/C ≧ 125, the test compound was         judged to have a remarkable antitumor effect in the dose.                

(2) Proliferation inhibition activity against mouse leukemia cell

Measurement method:

100 μl portions of a cell culturing medium (10% fetal bovineserum-containing-RPMI-1640 medium) containing 3×10³ mouse leukemia cell(P388) were put in a 96-hole microplate, the cells were cultured under5% CO₂ at 37° C. for 24 hours, 10 μl each of test solutions containingtest compounds respectively were added respectively, and the cells werefurther cultured under 5% CO₂ at 37° C. for 24 hours. 10 μl portions of0.5% Thiazoyl Blue were added to the culture broths, and incubation wascarried out under 5% CO₂ at 37° C. for 2 hours to carry out enzymaticreaction. 20% sodium dodecyl sulfate (SDS) was added to discontinue thereaction, incubation was further carried out at 37° C. for 4 hours todissolve the formed dye, and absorbance at 550 nm was measured andcompared with the control group. The results are shown in Table 3.

                  TABLE 3                                                         ______________________________________                                        Proliferation inhibition activity against mouse                               leukemia cell P388                                                                            50% inhibitory concentration                                  Test compound   (IC.sub.50, μM)                                            ______________________________________                                        Compound of Example 1                                                                         <0.030                                                        Compound of Example 2                                                                         0.29                                                          Compound of Example 3                                                                         0.065                                                         Compound of Example 4                                                                         0.096                                                         Compound of Example 5                                                                         0.28                                                          Compound of Example 6                                                                         0.059                                                         Compound of Example 7                                                                         0.091                                                         Compound of Example 8                                                                         0.30                                                          Compound of Example 9                                                                         0.028                                                         Compound of Example 10                                                                        0.46                                                          Compound of Example 11                                                                        <0.026                                                        Compound of Example 12                                                                        0.042                                                         Compound of Example 13                                                                        0.22                                                          Compound of Example 14                                                                        <0.027                                                        Compound of Example 15                                                                        0.31                                                          Compound of Example 17                                                                        0.044                                                         Compound of Example 22                                                                        0.11                                                          Compound of Example 23                                                                        <0.025                                                        Compound of Example 24                                                                        0.001                                                         Compound of Example 25                                                                        0.048                                                         Compound of Example 27                                                                        0.027                                                         Compound of Example 28                                                                        <0.029                                                        Compound of Example 29                                                                        0.005                                                         Compound of Example 30                                                                        0.003                                                         Compound of Example 31                                                                        0.011                                                         Compound of Example 33                                                                        0.11                                                          Compound of Example 34                                                                        0.019                                                         Compound of Example 35                                                                        0.17                                                          Compound of Example 36                                                                        0.002                                                         Compound of Example 37                                                                        0.095                                                         ______________________________________                                    

As apparent from the results of the above pharmacological test, thecompounds of this invention exhibit an excellent antitumor action, andare useful as an antitumor agent for control or prevention of diseases,particularly for treatment of cancers. When a compound of this inventionis used in such uses, it is, usually, formulated into a pharmaceuticalpreparation comprising an effective quantity of it and apharmaceutically acceptable carrier or diluent.

As administration forms at the time of use of a compound of thisinvention, various forms can be selected, and there can be mentionedoral agents such as, for example, tablet, capsules, powders, granules orliquids, or sterilized liquid parenteral agents such as, for examplesolutions or suspensions, or suppositories, ointments, or the like.

Solid preparations can be prepared, as they are, as forms of tablets,capsules, granules or powders, or can also be prepared using suitableadditives. Such additives may be additives usually used, and includesaccharides such as, for example, lactose and glucose; starches such as,for example, corn, wheat and rice; fatty acids such as, for example,stearic acid; inorganic salts such as, for example, magnesiummetasilicate aluminate and anhydrous calcium phosphate; synthesizedmacromolecules such as, for example, polyvinylpyrrolidone andpolyalkylene glycol; fatty acid salts such as, for example, calciumstearate and magnesium stearate; alcohols such as, for example, stearylalcohol and benzyl alcohol; synthesized cellulose derivatives such as,for example, methylcellulose, carboxymethylcellulose, ethylcellulose andhydroxypropylmethylcellulose; and further, gelatin, talc, vegetableoils, gum arabic, etc.

Solid preparations such as these tablets, capsules, granules and powderscontain an effective ingredient generally at 0.1-100 weight %,preferably at 5-100 weight %.

Liquid preparations are prepared in forms such as suspensions, syrups,injections or drops using suitable additives usually used in liquidpreparations such as water, alcohols or oils originated in vegetablessuch as, for example, soybean oil, peanut oil and sesame oil.

Particularly, solvents suitable in case of parenteral administration inthe form of intramuscular injection, intravenous injection orsubcutaneous injection include, for example, distilled water forinjection, aqueous lidocaine hydrochloride solutions (for intramuscularinjection), physiological saline, aqueous glucose solutions, ethanol,polyethylene glycol, liquids for intravenous injection (e.g. aqueoussolutions of citric acid and sodium citrate, etc.), electrolytesolutions (for intravenous drip and intravenous injection), etc., andtheir mixed solvents.

These injections can take forms that powder itself or to which suitableadditives were added is dissolved at the time of use, besides such formsthat ingredients are dissolved in advance. Such an injection containsusually 0.1-10 weight %, preferably 1-5 weight % of the effectiveingredient.

Further, a liquid agent of a suspension, syrup or the like for oraladministration can usually contain 0.5-10 weight % of the effectiveingredient.

The preferred dose of the compounds of this invention can be variedaccording to the kind of a compound to be used, the kind and applicationfrequency of the compounded composition, the specified site to betreated, the degree of diseases, the age of patients, diagnosis ofdoctors, the kind of tumor, etc., but, as an approximate standard, thedose per day and per one adult can, for example, be in the range of 10to 500 mg in case of oral administration, and in the range of 10 to 100mg in case of parenteral administration, preferably intravenousinjection. The administration frequency varies depending onadministration methods and symptoms, but is 1 to 5 times a day. Further,there can also be adopted administration methods such as intermittentadministration, e.g. every second day administration or every third dayadministration.

This invention is more specifically described below by examples, but notlimited only by these examples.

EXAMPLE A

The compound represented by the formula ##STR16##

3.4 g of12,13-dihydro-1,11-dihydroxy-13-(β-D-glucopyranosyl)-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dionewas dissolved in 120 ml of 10% aqueous potassium hydroxide solution, andthe solution was stirred at room temperature for 2 hours. The reactionsolution was neutralized with addition of 120 ml of 2N hydrochloricacid, and the precipitated red crystals were filtered, washed with waterand dried to give 3.0 g of the captioned compound.

FAB-MS(m/z): 520 (M)⁺, 521 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 3.42 (1H,m), 3.56-3.70 (2H,m), 3.76(1H,m), 3.95-4.10 (2H,m), 4.95 (1H,d,J=4.6 Hz), 5.24 (1H,d,J=5.4 Hz),5.32 (1H,dd,J=4.9, 5.1 Hz), 7.06 (2H,dd,J=7.6, 7.8 Hz), 7.09 (1H,d,J=8.0Hz), 7.20 (1H,d,J=7.8 Hz), 7.40 (1H,d,J=7.8 Hz), 8.36 (1H,d,J=7.6Hz),8.51 (1H,d,J=7.6 Hz), 10.13 (1H,s), 10.52 (1H,s), 11.11 (1H,s)

EXAMPLE B

The compound represented by the formula ##STR17##

50 mg of rebeccamycin was dissolved in 5 ml of N,N-dimethylformamide, 5ml of 2N aqueous sodium hydroxide solution was added, and the mixturewas stirred at 80° C. for 3 hours. 60 ml of water was added to thereaction solution, the mixture was cooled with ice, and the precipitatedyellow precipitate was recovered by filtration. This was subjected tocolumn chromatography on silica gel (inner diameter 1.5 cm, length 45cm), the column was washed with chloroform, elution was carried out withchloroform-tetrahydrofuran (10:1), and the fraction containing thedesired product was concentrated to dryness. The resultant yellow powderwas washed with chloroform to give 6.4 mg of the captioned compound.

Rf value: 0.51 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran:acetic acid=10:1:1:0.2)

FAB-MS(m/z): 571 [M+H]⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.9 (1H,s), 9.07 (1H,d,J=7.8 Hz),8.92 (1H,d,J=7.8 Hz), 7.78 (2H,t,J=7.8 Hz), 7.53 (1H,d,J=7.8 Hz), 7.50(1H,d,J=7.8, Hz), 7.03 (1H,d,J=8.9 Hz), 3.96 (2H,m), 3.87 (1H,m), 3.61(3H,s), 3.54-3.73 (3H,m).

EXAMPLE C

The compound represented by the formula ##STR18##

1.3 g of12,13-dihydro-2,10-dihydroxy-13-(β-D-glucopyranosyl)-6-methyl-5H-indolo(2,3-a)pyrrolo(3,4-c)carbazole-5,7(6H)-dionewas dissolved in 40 ml of 10% aqueous potassium hydroxide solution, andthe solution was stirred at room temperature for 1 hour. 40 ml of 2Nhydrochloric acid was added to the reaction solution to neutralize it,and the mixture was extracted with 1 L of methyl ethyl ketone. Theorganic layer was washed with saturated saline, dried, and concentrated.The residue was recrystallized from acetone-heptane to give 1.2 g of thecaptioned compound.

Rf value: 0.40 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; toluene:acetonitrile:tetrahydrofuran:water:aceticacid=2:4:2:0.5:0.1)

FAB-MS (m/z): 520 (M)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.4 (1H, s), 9.95 (1H, s), 9.92(1H, s), 8.69 (1H, d, J=7.7 Hz), 8.63 (1H, d, J=7.7 Hz), 7.25 (1H, d,J=1.5 Hz), 7.03 (1H, d, J=1.5 Hz), 6.90 (1H, dd, J=7.7 Hz, 1.5 Hz), 6.87(1H, dd, J=7.7 Hz, 1.5 Hz). 6.06 (1H, d, 8.0 Hz), 5.95 (1H, t, J=4.6Hz), 5.38 (1H, d, J=5.1 Hz), 5.16 (1H, d, J=5.2 Hz), 4.99 (1H, d, J=5.2Hz), 3.30-4.10 (6H, m).

EXAMPLE 1

The compound represented by the formula ##STR19##

3.51 g of12,13-dihydro-1,11-dihydroxy-13-(β-D-glucopyranosyl)-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dionewas dissolved in 8 ml of hydrazine hydrate (Wako Pure ChemicalIndustries, Ltd.), and reaction was carried out at room temperature for2 hours. After the reaction, 180 ml of purified water was added, the pHof the solution was adjusted to 5.0 with concentrated hydrochloric acid,the mixture was sufficiently cooled with ice, and the resultingprecipitate was collected by filtration, washed with purified water anddried under reduced pressure to give 3.51 g of the captioned compoundrepresented by the formula (1). (yield: 97%)

FAB-MS (m/z): 535 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, brs), 10.4 (1H, s), 10.0(1H, s), 8.72 (1H, d, J=7.8 Hz), 8.54 (1H, d, J=7.8 Hz), 7.19 (2H, t,J=7.8 Hz). 7.19 (2H, t, J=7.8 Hz), 7.05 (1H, d, J=9.3 Hz), 7.02 (1H, d,J=7.8 Hz), 7.00 (1H, d, J=7.8 Hz), 5.42 (1H, brd, J=5.8 Hz), 5.35 (1H,brs), 5.22 (1H, brd, J=4.4 Hz), 4.96 (2H, brs), 4.91 (1H, brd, J=5.3Hz), 4.01 (2H, m), 3.73 (1H, m), 3.63 (2H, m), 3.39 (1H, m)

EXAMPLE 2

The compound represented by the formula ##STR20##

3.47 g of the compound obtained in Example 1 was dissolved in 20 ml ofN,N-dimethylformamide (DMF), while the solution was stirred at roomtemperature, 20 ml of a 100 mg/ml solution of glyoxylic acid (Sigma Co.)was added portionwise, and thereby a precipitate was formed andsolidified into a gel-like state. Further, 200 ml of purified water wasadded, the reaction solution was cooled with ice, and the resultantprecipitate was collected by filtration, washed with purified water anddried under reduced pressure to give 3.85 g of the captioned compoundrepresented by the formula (2), (yield: 100%)

FAB-MS (m/z): 591 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.1 (1H, brs), 10.5 (1H, brs),10.1 (1H, brs), 9.01 (1H, s), 8.69 (1H, d, J=7.8 Hz), 8.53 (1H, d, J=7.8Hz), 7.23 (2H, t, J=7.8 Hz), 7.10 (1H, d, J=9.3 Hz), 7.06 (1H, d, J=7.8Hz), 7.04 (1H, d, J=7.8 Hz), 5.44 (1H, brs), 5.34 (1H, brs), 5.24 (1H,brs), 4.95 (1H, brd, J=5.9 Hz), 4.02 (2H, m), 3.76 (1H, m), 3.64 (2H,m), 3.40 (1H, m)

EXAMPLE 3

The Compound represented by the formula ##STR21##

24 mg of the compound obtained in Example 1 was dissolved in 0.5 ml ofN,N-dimethylformamide (DMF), while the solution was stirred at roomtemperature, 0.2 ml of 15% succinic semialdehyde (Aldrich Chemical Co.)was added, and one hour later, 5 ml of purified water was added. Afterthe reaction solution was cooled with ice, and the resultant precipitatewas collected by filtration, washed with purified water and dried underreduced pressure to give 25.3 mg of the captioned compound representedby the formula (3). (yield: 91%)

FAB-MS (m/z): 619 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 12.1 (1H, brs), 11.0 (1H, brs),10.4 (1H, brs), 10.0 (1H, brs), 8.69 (1H, brs), 8.68 (1H, d, J=7.8 Hz),8.51 (1H, d, J=8.3 Hz), 7.19 (2H, t, J=7.8 Hz), 7.07 (1H, d, J=9.3 Hz),7.04 (1H, d, J=7.8 Hz), 7.01 (1H, d, J=7.8 Hz), 5.43 (1H, brd, J=5.4Hz), 5.33 (1H, brs), 5.22 (1H, brs), 4.93 (1H, brd, J=4.9 Hz), 4.01 (2H,m), 3.74 (1H, m), 3.63 (2H, m), 3.40 (1H, m)

EXAMPLE 4

The compound represented by the formula ##STR22##

511 mg of rebeccamycin [the compound described in J. Antibiotics 40,668-678 (1987)] was dissolved in 3 ml of hydrazine hydrate (Wako PureChemical Industries, Ltd.), and the solution was allowed to stand atroom temperature for one hour. 200 ml of purified water was added, andthe resultant precipitate was collected by filtration, washed with 100ml of purified water and dried under reduced pressure to give 497 mg ofthe captioned 6-N-aminorebeccamycin represented by the formula (4).(yield: 95%)

FAB-MS (m/z): 585 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.64 (1H, brs), 9.24 (1H, d, J=7.8Hz), 9.07 (1H, d, J=7.8 Hz), 7.70 (2H, t, J=7.8 Hz), 7.45 (1H, d, J=7.8Hz), 7.42 (1H, d, J=7.8 Hz), 6.93 (1H, d, J=8.8 Hz), 5.42 (1H, d, J=5. 8Hz), 5.33 (1H, t, J=5.4 Hz), 5.03 (3H, brs), 3.97 (2H, m), 3.84 (1H, m),3.59 (3H, s), 3.50˜3.70 (3H, m)

EXAMPLE 5

The compound represented by the formula ##STR23## [Process A]

5 g of the compound obtained in Example 1 was dissolved in 60 ml ofN,N-dimethylformamide, 1.8 ml of concentrated hydrochloric acid wasadded, the mixture was heated at 60° C. for 4 hours, 0.8 ml ofconcentrated hydrochloric acid was further added, and the mixture waswarmed at 37° C. for 16 hours. This was mixed with 1 l of ethyl acetate,the mixture was washed successively with 2% sodium bicarbonate aqueoussolution and water, and then the ethyl acetate layer was dehydrated withanhydrous sodium sulfate and concentrated to dryness to give 3.3 g oforange powder. This was dissolved in methanol and subjected to columnchromatography on Sephadex LH 20 (inner diameter 3 cm, length 54 cm,eluted with methanol), and the fractions containing the desired productwere concentrated to dryness to give 2413.6 mg of the captioned compoundrepresented by the formula (5) as orange powder.

[Process B]

25.9 mg of the compound obtained in Example A was dissolved in 0.5 ml ofN,N-dimethylformamide, 15.0 mg of formohydrazide was added, and themixture was stirred at 70° C. for 2 hours. This was mixed with 70 ml ofethyl acetate, and the mixture was washed with water (20 ml). The ethylacetate layer was dehydrated with anhydrous sodium sulfate andconcentrated to dryness to give 26.9 mg of orange powder. This wasdissolved in methanol and subjected to column chromatography on SephadexLH 20 (inner diameter 1.5 cm, length 48 cm, eluted with methanol), andthe fractions containing the desired product were concentrated todryness to give 16.3 mg of the captioned compound represented by theformula (5) as orange powder.

Rf value: 0.35 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform-methanol-tetrahydrofuran=2:1:1)

FAB-MS (m/z): 562 (M)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, brs), 10.8 (1H, s), 10.4(1H, s), 10.0 (1H, s), 8.64 (1H, d, J=8.3 Hz), 8.47 (1H, d, J=8.3 Hz),8.44 (1H, s), 7.21 (2H, t, J=7.8 Hz), 7.06 (1H, d, J=9.7 Hz), 7.05 (1H,d, J=7.8 Hz), 7.02 (1H, d, J=7.8 Hz), 5.43 (1H, d, J=5.8 Hz), 5.36 (1H,brs), 5.22 (1H, d, J=5.4 Hz), 4.92 (1H, d, J=5.4 Hz), 4.02 (2H, m), 3.75(1H, m), 3.62 (2H, m), 3.39 (1H, m)

EXAMPLE 6

The compound represented by the formula ##STR24##

30 ml of acetic acid and 2 ml of acetic anhydride were added to 510 mgof the compound obtained in Example 1, and the compound was dissolvedtherein with heating at 90° C. Water was added thereto to make themixture 300 ml, and the reaction product was adsorbed on a column ofDiaion HP 20 (inner diameter 3 cm, length 13.5 cm) and after washing thecolumn with 600 ml of water, eluted with 300 ml of methanol. Themethanol eluate was concentrated to dryness, the residue was dissolvedin 50 ml of methanol, and the solution was concentrated to about 5 ml.100 ml of ethyl acetate was added thereto, the mixture was allowed tostand overnight at 4° C., and the resultant orange precipitate wascollected by filtration to give 426 mg of the captioned compoundrepresented by the formula (6).

Rf value: 0.43 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform-methanol-tetrahydrofuran=2:1:1)

FAB-MS (m/z): 576 (M)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.7 (1H, s), 10.4(1H, brs), 10.05 (1H, s), 8.64 (1H, d, J=7.8 Hz), 8.47 (1H, d, J=7.8Hz), 7.20 (2H, t, J=7.8 Hz), 7.01-7.06 (3H, m), 5.35-5.45 (2H, m), 5.23(1H, brs), 4.92 (1H, brs), 4.02 (2H, m), 3.74 (1H, m), 3.58-3.70 (2H,m), 3.40 (1H, m), 2.10 (3H, s)

EXAMPLE 7

The compound represented by the formula ##STR25##

72.5 mg of the compound obtained in Example 1 was dissolved in a mixtureof 8 ml of tetrahydrofuran and 5 ml of methanol, 140 μl of 2Nhydrochloric acid and 13.2 μl of 37% formaldehyde aqueous solution wereadded, and the mixture was stirred at room temperature for 2 hours andconcentrated to dryness. This was dissolved in 5 ml ofN,N-dimethylformamide, 80 mg of 10% palladium carbon, and the mixturewas subjected to reduction under hydrogen gas at room temperature for 2hours and then filtered on Celite. 80 ml of ethyl acetate was added tothe resultant filtrate, the mixture was washed successively with 2%sodium bicarbonate aqueous solution and water, and then the resultantethyl acetate layer was dehydrated and concentrated to dryness to give28.8 mg of orange powder. This was dissolved in a small quantity ofmethanol, and the solution was subjected to column chromatography onSephadex LH-20 (inner diameter 1.5 cm, length 90 cm, eluted withmethanol) to give 17.1 mg of the captioned compound represented by theformula (7) as orange powder.

Rf value: 0.49 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform-methanol-tetrahydrofuran-acetic acid=20:10:10:1)

FAB-MS (m/z): 549 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, s), 10.4 (1H, s), 9.98(1H, s), 8.72 (1H, d, J=7.8 Hz), 8.54 (1H, d, J=7.8 Hz), 7.19 (2H, t,J=7.8 Hz), 7.00-7.06 (3H, m), 5.73 (1H, q, J=5.4 Hz), 5.43 (1H, d, J=5.7Hz), 5.35 (1H, brs), 5.22 (1H, d, J=5.4 Hz), 4.90 (1H, d, J=5.4 Hz),3.96-4.03 (2H, m), 3.74 (1H, m), 3.58-3.70 (2H, m), 3.40 (1H, m), 2.74(3H, d, J=5.4 Hz)

EXAMPLE 8

The compound represented by the formula ##STR26##

500 mg of the compound obtained in Example 2 was dissolved in 6 ml ofN,N-dimethylformamide(DMF), 75 mg of 10% palladium-carbon (Pd-C) wasadded, and hydrogenation was carried out at room temperature for 3.5hours under the stirring. The reaction mixture was filtered using filterpaper on which diatom earth was spread to remove Pd-C, and 150 ml ofwater was added to the filtrate. The mixture was adjusted to pH 5 with1N NaOH, and then extracted with ethyl acetate (200 ml×5). The ethylacetate layer was concentrated, and the precipitated crystals werecollected by filtration to give 182.3 mg of the captioned compoundrepresented by the formula (8).

FAB-MS (m/z): 593 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 12.6 (1H, brs), 10.9 (1H, s), 10.4(1H, s), 10.0 (1H, s), 8.69 (1H, d, J=8.3 Hz), 8.52 (1H, d, J=7.8 Hz),7.18 (2H, t, J=7.8 Hz), 6.99˜7.05 (3H, m), 5.90 (1H, brs), 5.42 (1H, d,J=5.4 Hz), 5.35 (1H, t, J=5.4 Hz), 5.21 (1H, d, J=4.9 Hz), 4.89 (1H, d,J=5.4 Hz), 4.03 (2H, m), 3.83 (2H, s), 3.74 (1H, m), 3.63 (2H, m), 3.39(1H, m)

EXAMPLE 9

The compound represented by the formula ##STR27##

30 ml of methanol was added to 501.7 mg of the compound obtained inExample A and 501.7 mg of semicarbazide hydrochloride, 0.325 ml oftriethylamine was then added, and the mixture was refluxed with heatingfor 8 hours. After the reaction, the reaction solution was concentratedto dryness, 300 ml of methyl ethyl ketone (MEK) and 200 ml of water wereadded, extraction operation was carried out, and another 300 ml of MEKwas added to the water layer to carry out reextration. The MEK layerswere combined and concentrated to dryness, 300 ml of methanol was addedto the residue to dissolve it, the solution was subjected to achromatograph tower of Sephadex LH-20 (3×28 cm), and elution was carriedout with methanol. The fractions containing the desired product wereconcentrated to dryness to give 461 mg of the captioned compoundrepresented by the formula (g) as red crystal-like powder.

Rf value: 0.15 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 577 (M)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.4 (1H, s), 10.0(1H, s), 8.68 (1H, d, J=7.8 Hz), 8.66 (1H, brs), 8.51 (1H, d, J=7.8 Hz),7.20 (2H, t, J=7.8 Hz), 7.01˜7.07 (3H, m), 6.41 (2H, brs), 5.44 (1H, d,J=5.4 Hz), 5.38 (1H, brs), 5.23 (1H, d, J=4.9 Hz), 4.91 (1H, brs),4.00˜4.09 (2H, m), 3.75 (1H, m), 3.60˜3.68 (2H, m), 3.39 (1H, m)

EXAMPLE 10

The compound represented by the formula ##STR28##

4 ml of methanol was added to 22 mg of the compound obtained in ExampleA and 20 mg of thiosemicarbazide, and the mixture was refluxed withheating for 22 hours. The reaction solution was concentrated to dryness,the residue was dissolved in 4 ml of methanol, the solution wassubjected to a chromatograph tower of Sephadex LH-20 (1.8×35 cm), andelution was carried out with methanol. The fractions containing thedesired product were concentrated to dryness to give 10.7 mg of thecaptioned compound represented by the formula (10).

Rf value: 0.29 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 594 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.4 (1H, brs), 10.1(1H, brs), 9.73 (1H, brs), 8.65 (1H, d, J=7.8 Hz), 8.49 (1H, d, J=7.8Hz), 8.27 (2H, s), 7.21 (2H, t, J=7.8 Hz), 7.01˜7.12 (3H, m), 5.45 (1H,brs), 5.37 (1H, brs), 5.24 (1H, brs), 4.91 (1H, brs), 3.97˜4.10 (2H, m),3.74 (1H, m), 3.62 (2H, m), 3.40 (1H, m)

EXAMPLE 11

The compound represented by the formula ##STR29##

9.5 mg of the compound obtained in Example 1 was dissolved in 2 ml oftetrahydrofuran (THF), to the solution was added 30 mg ofmethanesulfonic anhydride (Aldrich Chemical Co.), and the mixture wasallowed to stand at room temperature for 48 hours. The reaction solutionwas concentrated to dryness, and the residue was dissolved in 2 ml ofmethanol, subjected to a chromatograph tower of Sephadex LH-20 (1.8×34cm), and eluted with methanol. The fractions containing the desiredproduct were concentrated to dryness to give 8.3 mg of the captionedcompound represented by the formula (11).

Rf value: 0.48 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 612 (M)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.5 (1H, brs), 10.4(1H, s), 10.1 (1H, s), 8.67 (1H, d, J=7.9 Hz), 8.50 (1H, d, J=7.7 Hz),7.22 (2H, t, J=7.6 Hz), 7.02˜7.07 (3H, m), 5.43 (1H, d, J=5.8 Hz), 5.36(1H, brs), 5.22 (1H, d, J=5.2 Hz), 4.89 (1H, d, J=4.8 Hz), 4.03 (2H, m),3.75 (1H, m), 3.63 (2H, m), 3.40 (1H, m)

EXAMPLE 12

The compound represented by the formula ##STR30##

1 ml of the methanol and 2 ml of tetrahydrofuran were added to 11.7 mgof the compound obtained in Example 1 to make a solution, 0.1 ml ofpropionic anhydride (Aldrich Chemical Co.) was added, and the mixturewas stirred at room temperature for 4 hours. 2 ml of water and 3 ml ofmethanol were added to the reaction solution, the mixture was allowed tostand for 30 minutes and then concentrated to dryness, and 3 ml ofmethanol was added to make a solution. The solution was subjected to achromatograph tower of Sephadex LH-20 (1.8×30 cm) and eluted withmethanol, and the fractions containing the desired product wereconcentrated to dryness to give 6.2 mg of the captioned compoundrepresented by the formula (12).

Rf value: 0.55 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.6 (1H, brs), 10.4(1H, brs), 10.0 (1H, s), 8.64 (1H, d, J=7.8 Hz), 8.47 (1H, d, J=7.8 Hz),7.20 (2H, t, J=7.8 Hz), 7.00˜7.08 (3H, m), 5.30˜5.45 (2H, m), 5.21 (1H,m), 4.92 (1H, m), 4.02 (2H, m), 3.75 (1H, m), 3.62 (2H, m), 3.38 (1H,m), 2.39 (2H, q, J=9.3 Hz), 1.16 (3H, t, J=7.3 Hz)

EXAMPLE 13

The compound represented by the formula ##STR31##

9.9 mg of the compound obtained in Example 1 was dissolved in 2 ml oftetrahydrofuran, 0.06 ml of trifluoroacetic anhydride (Aldrich ChemicalCo.) was added, and the mixture was allowed to stand at room temperaturefor 15 minutes. 2 ml of water was added to the reaction solution, themixture was concentrated to dryness, 2 ml of water and 10 ml of ethylacetate were added, extraction operation was carried out, and theresultant ethyl acetate layer was concentrated to dryness. The resultantcrude substance was dissolved in 3 ml of methanol, subjected to achromatograph tower of Sephadex LH-20 (1.8×30 cm) and eluted withmethanol, and the fractions containing the desired product wereconcentrated to dryness to give 9.5 mg of the captioned compoundrepresented by the formula (13).

Rf value: 0.53 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z) : 630 (M)⁺

¹ H-NMR (500 MHz, DMSO-d₆), δ (ppm): 12.7 (1H, brs), 11.0 (1H, brs),10.5 (1H, brs), 10.1 (1H, brs), 8.61 (1H, d, J=7.6 Hz), 8.45 (1H, d,J=7.9 Hz), 7.21 (2H, t, J=7.6 Hz), 7.02˜7.07 (3H, m), 5.42 (1H, d, J=5.8Hz), 5.35 (1H, brs), 5.21 (1H, brs), 4.91 (1H, d, J=5.5 Hz), 4.02 (2H,m), 3.76 (1H, m), 3.61 (2H, m), 3.39 (1H, m)

EXAMPLE 14

The compound represented by the formula ##STR32##

4 ml of methanol and 4 ml of benzene were added to 31.6 mg of thecompound obtained in Example 8 to make a solution, 0.15 ml oftrimethylsilyldiazomethane (10% hexane solution, Tokyo Kaset Co.) wasadded, and the mixture was allowed to stand at room for 10 minutes andconcentrated to dryness to give 29.3 mg of the methyl ester of thecompound obtained in Example 8. This was dissolved in 5 ml of methanol,0.6 ml of concentrated ammonia water was added, and the mixture wasstirred at room temperature for 16 hours. The reaction solution wasconcentrated to dryness, 3 ml of methanol was added to the residue tomake a solution, and the solution was subjected to a chromatograph towerof Sephadex LH-20 (1.8×36 cm) and eluted with methanol. The fractionscontaining the desired product were concentrated to dryness to give 16.9mg of the captioned compound represented by the formula (14).

Rf value: 0.22 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 592 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, s), 10.4 (1H, brs), 10.0(1H, brs). 8.69 (1H, d, J=7.3 Hz), 8.52 (1H, d, J=8.3 Hz), 7.77 (1H,brs), 7.39 (1H, brs), 7.19 (2H, t, J=7.8 Hz), 6.98˜7.05 (3H, m), 6.25(1H, t, J=3.9 Hz), 5.41 (1H, d, J=5.4 Hz), 5.35 (1H, brs), 5.20 (1H, d,J=5.4 Hz), 4.87 (1H, d, J=5.4 Hz), 4.02 (2H, m), 3.74 (1H, m), 3.68˜3.70(4H, m), 3.39 (1H, m)

EXAMPLE 15 ##STR33##

2 ml of methanol was added to 11 mg of the compound obtained in ExampleA and 10 mg of 2-hydrazinopyridine (Aldrich Chemical Co.) to make asolution, and the solution was refluxed with heating for 1.5 hours. Thereaction solution was concentrated to dryness, 30 ml of water and 50 mlof ethyl acetate were added, the water layer was adjusted to pH 5 with1N hydrochloric acid, extraction operation was carried out, and theresultant ethyl acetate layer was concentrated to dryness. The resultantcrude substance was dissolved in 2 ml of methanol, subjected to achromatograph tower of Sephadex LH-20 (1.8×36 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 10 mg of the captioned compound represented by theformula (15).

Rf value: 0.46 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 612 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.4 (1H, s), 10.0(1H, s), 9.34 (1H, s), 8.65 (1H, d. J=8.3 Hz), 8.48 (1H, d, J=7.8 Hz),7.95 (1H, d, J=4.9 Hz), 7.62 (1H, t, J=7.8 Hz), 7.18 (2H, t, J=7.8 Hz),7.00˜7.08 (3H, m), 6.86 (1H, d, J=7.8 Hz), 6.78 (1H, dd, J=4.9, 7.8 Hz),5.44 (1H, d, J=5.8 Hz), 5.37 (1H, brs), 5.23 (1H, d, J=5.8 Hz), 4.92(1H, brs), 4.02 (2H, m), 3.76 (1H, m), 3.64 (2H, m), 3.41 (1H, m)

EXAMPLE 16

The compound represented by the formula ##STR34##

4 ml of methanol was added to 24 mg of the compound obtained in ExampleA and 4-hydrazinobenzoic acid (Aldrich Chemical Co.), and the mixturewas refluxed with heating for 2 hours. The reaction solution wassubjected to a chromatograph tower of Sephadex LH-20 (1.8×44 cm) andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness to give 20.9 mg of the captioned compoundrepresented by the formula (16) as red crystal-like powder.

Rf value: 0.31 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 655 (M+H)⁺

¹ H-NMR (500 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.5 (1H, brs), 10.1(1H, brs), 9.11 (1H, s), 8.65 (1H, d, J=7.9 Hz), 8.48 (1H, d, J=7.9 Hz),7.80 (2H, d, J=8.3 Hz), 7.18 (2H, t, J=7.6 Hz), 7.01˜7.08 (3H, m), 6.84(2H, d, J=8.3 Hz), 5.20˜5.60 (3H, brs), 4.96 (1H, brs), 4.03 (2H, m),3.76 (1H, m), 3.65 (2H, m), 3.41 (1H, m)

EXAMPLE 17

The compound represented by the formula ##STR35##

6 ml of 50% methanol was added to 26 mg of the compound obtained inExample A and 38 mg of oxamic hydrazine (Aldrich Chemical Co.), and themixture was stirred with heating at 80° C. for 20 hours. The reactionsolution was concentrated to dryness, 15 ml of water and 50 ml of ethylacetate were added, the mixture was adjusted to pH 2 with 1Nhydrochloric acid, and extraction operation was carried out. The ethylacetate layer was concentrated, and the precipitated crystals werecollected by filtration to give 10 mg of the captioned compoundrepresented by the formula (17).

Rf value: 0.38 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 606 (M+H)⁺

¹ H-NMR (500 MHz, DMSO-d₆), δ (ppm): 11.4 (1H, s), 11.0 (1H, s), 10.4(1H, s), 10.0 (1H, s), 8.63 (1H, d, J=7.9 Hz), 8.46 (1H, d, J=7.9 Hz),8.38 (1H, s), 8.11 (1H, s), 7.21 (2H, t, J=7.9 Hz), 7.02˜7.07 (3H, m),5.41 (1H, d, J=5.8 Hz), 5.35 (1H, t, J=5.8 Hz), 5.19 (1H, d, J=5.2 Hz),4.89 (1H, d, J=5.5 Hz), 4.03 (2H, m), 3.76 (1H, m), 3.63 (2H, m), 3.40(1H, m)

EXAMPLE 18

The compound represented by the formula ##STR36##

26.7 mg of the compound obtained in Example 1 and 5.5 mg of succinicanhydride were dissolved in 0.5 ml of pyridine, and the solution wasstirred at room temperature for 18 hours. This was concentrated todryness under reduced pressure and the residue was dissolved in a smallquantity of N,N-dimethylformamide and subjected to high performanceliquid chromatography (HPLC) [Chromatolex ODS, 20×250 mm, moving phase:20% acetonitrile]. The fractions containing the desired product wereconcentrated to remove acetonitrile, adjusted to pH 2 and extracted with100 ml of ethyl acetate. The ethyl acetate layer was dehydrated withanhydrous sodium sulfate and concentrated to dryness. The residue wasdissolved in methanol and subjected to column chromatography on SephadexLH-20 (inner diameter 1.5 cm, length 90 cm, eluted with methanol), andthe fractions containing the desired product were concentrated todryness to give 9.7 mg of the captioned compound represented by theformula (18) as orange powder.

HPLC; Rt, 5.3 minutes (column: Chromatolex ODS, inner diameter 4.6 mm,length 250 mm, detection; UV 305 nm, flow rate; 1 ml/minute, movingphase; 27.5% acetonitrile:trifluoroacetic acid=1000:1)

FAB-MS (m/z): 657 (M+Na)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.0 (1H, s), 10.7 (1H, brs), 10.4(1H, brs), 10.1 (1H, brs), 8.64 (1H, d, J=7.9 Hz), 8.47 (1H, d, J=7.9Hz), 7.19 (2H, t, J=7.8 Hz), 7.01˜7.07 (3H, m), 5.42 (2H, brs), 5.22(1H, brs), 4.92 (1H, brs), 4.02 (2H, m), 3.75 (1H, m), 3.63 (2H, m),3.40 (1H, m), 2.65 (2H, t, J=7.3 Hz), 2.52 (2H, t, J=7.3 Hz)

EXAMPLE 19

The compound represented by the formula ##STR37##

30 mg of the compound obtained in Example 1 was dissolved in 0.5 ml ofN,N-dimethylformamide, 0.1 ml of methyl iodide was added, and themixture was stirred at room temperature for 18 hours. This was mixedwith 50 ml of ethyl acetate, the mixture was washed successively with 1%sodium bicarbonate aqueous solution and then water, and the ethylacetate layer was dehydrated with anhydrous sodium sulfate andconcentrated to dryness. The residue was dissolved in methanol andsubjected to column chromatography on Sephadex LH-20 (1.5×90 cm, elutedwith methanol), and the fractions containing the desired product wereconcentrated to dryness to give 18.0 mg of the captioned compoundrepresented by the formula (19) as orange powder.

Rf value: 0.51 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran:acetic acid=20:10:10:1)

FAB-MS (m/z): 563 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, s), 10.3 (1H, s), 9.95(1H, s), 8.70 (1H, d, J=8.3 Hz), 8.53 (1H, d, J=8.3 Hz), 7.18 (2H, t,J=7.8 Hz), 7.00˜7.06 (3H, m), 5.41 (1H, d, J=5.4 Hz), 5.34 (1H, t, J=5.4Hz), 5.19 (1H, d, J=5.4 Hz), 4.86 (1H, d, J=5.4 Hz), 4.02 (2H, m), 3.75(1H, m), 3.62 (2H, m), 3.39 (1H, m), 3.02 (6H, s)

EXAMPLE 20

The compound represented by the formula ##STR38##

82.1 mg of t-butyloxycarbonyl (Boc)-glycine was dissolved in 1 ml ofmethylene chloride, the solution was stirred under ice cooling for 15minutes, 96.7 mg of dicyclohexylcarbodiimide dissolved in 1 ml ofmethylene chloride was added, and the mixture was stirred under icecooling for 15 minutes. To this was added 227.6 mg of the compoundobtained in Example 1 dissolved in 6 ml of pyridine, and the mixture wasstirred at room temperature for 17 hours. The reaction solution wasconcentrated to dryness, the residue was dissolved in ethyl acetate, thesolution was washed successively with saturated saline, acidic water (pH2) and then water, and the ethyl acetate layer was dehydrated withanhydrous sodium sulfate and concentrated to dryness. The residue wassubjected to silica gel column chromatography (1.5×55 cm, eluted withtoluene:methanol=6:1), and the fractions containing the desired productwere concentrated to dryness to give 105.2 mg of the Boc derivative ofthe captioned compound represented by the formula (20) as orange powder.This was dissolved in 1.2 ml of trifluoroacetic acid, and the solutionwas stirred at room temperature for 30 minutes to remove the Boc group.The reaction solution was concentrated to dryness, the residue wasdissolved in 15 ml of water, and the solution was adjusted to pH 7.5-8and extracted with n-butanol. 40 ml of water was added to the n-butanollayer (50 ml), and the mixture was adjusted to pH 2 with dilutehydrochloric acid and concentrated to dryness. The resultant orangepowder was dissolved in methanol and subjected to column chromatographyon Sephadex LH-20 (1.5×38 cm, eluted with methanol), and the fractionscontaining the desired product were concentrated to dryness to give 63.7mg of the hydrochloride of the captioned compound represented by theformula (20) as orange powder.

HPLC; Rt, 8.7 minutes (column: Chromatolex ODS, inner diameter 4.6 mm,length 250 mm, detection; UV 305 nm, flow rate; 1 ml/minute, movingphase; 20% acetonitrile:trifluoroacetic acid=1000:1→70%acetonitrile:trifluoroacetic acid=1000:1, 30 minutes linear gradient)

FAB-MS(m/z): 592 (M+H)⁺

¹ H-NMR (hydrochloride, 400 MHz, DMSO-d₆), δ (ppm): 11.3 (1H, brs), 11.0(1H, brs), 10.5 (1H, s), 10.1 (1H, s), 8.62 (1H, d, J=8.3 Hz). 8.46 (1H,d, J=8.3 Hz), 8.31 (2H, s), 7.19 (2H, t, J=7.8 Hz), 7.03˜7.08 (3H, m),5.46 (1H, brs), 5.34 (1H, brs), 5.27 (1H, brs), 4.91 (1H, brd, J=4.9Hz), 4.03 (2H, m), 3.98 (2H, s), 3.76 (1H, m), 3.64 (2H, m), 3.40 (1H,m)

EXAMPLE 21

The compound represented by the formula ##STR39##

40.0 mg of the compound obtained in Example A was dissolved in 3 ml ofN,N-dimethylformamide, 42.2 mg of 1-deoxy-1-hydrazino-D-sorbitol and 0.1ml of triethylamine was added, and the mixture was refluxed with heatingfor 16 hours. This was brought back to room temperature, subjected to achromatograph tower of Sephadex LH-20 (1.8×20 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 20.0 mg of the captioned compound represented by theformula (21).

FAB-MS (m/z): 699 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.91 (1H, s), 10.35 (1H, brs),9.96 (1H, brs), 8.73 (1H, d, J=8.9 Hz), 8.54 (1H, d, J=8.9 Hz), 7.20(2H, t, J=8.4 Hz), 7.00-7.10 (3H, m), 5.76 (1H, t, J=3.8 Hz), 5.42 (1H,d, J=5.5 Hz), 5.37 (1H, brs), 5.22 (1H, d, J=5.5 Hz), 4.89 (1H, brs),4.67 (1H, d, J=3.4 Hz), 4.45 (1H, d, J=5.1 Hz), 4.37 (1H, d, J=7.0 Hz),4.25-4.43 (2H, m), 4.00 (2H, m), 3.55-3.80 (7H, m), 3.44-3.52 (2H, m),3.35-3.44 (2H, m), 3.05-3.20 (2H, m)

EXAMPLE 22

The compound represented by the formula ##STR40##

100 mg of the compound obtained in Example A was dissolved in 5 ml ofN,N-dimethylformamide, 100 mg of carbohydrazide was added, and themixture was stirred at 80° C. for 3 hours and concentrated to dryness.The residue was dissolved in methanol and the insoluble matters wereremoved by Celite filtration. The resultant filtrate was concentrated,and the residue was dissolved in a small quantity of methanol, subjectedto a chromatograph tower of Sephadex LH-20 (1.5×20 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 91.2 mg of the captioned compound represented by theformula (22).

Rf value: 0.1 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform-methanol-tetrahydrofuran=2:1:1:)

FAB-MS (m/z): 593 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.96 (1H, s), 10.40 (1H, s), 10.01(1H, s), 8.95 (1H, s), 8.65 (1H, d, J=8.2 Hz), 8.50 (1H, d, J=8.2 Hz),7.90 (1H, s), 7.17 (2H, t, J=6.9 Hz), 7.00-7.10 (3H, m), 5.43 (1H, d,J=4.1 Hz), 5.38 (1H, brs), 5.20 (1H, s), 4.90 (1H, s), 4.39 (2H, brs),4.04 (2H, m), 3.75 (1H, m), 3.55-3.70 (2H, m), 3.38 (1H, m)

EXAMPLE 23

The compound represented by the formula ##STR41##

15.0 mg of the compound obtained in Example A was dissolved in 1 ml ofN,N-dimethylformamide, 32 mg of 3-hydoxybenzylhydrazine dihydrochlorideand 0.1 ml of 10% sodium bicarbonate aqueous solution were added, andthe mixture was stirred at 80° C. for 4 hours. This was mixed with 50 mlof ethyl acetate, the mixture was washed successively with 0.2Nhydrochloric acid and then saturated saline, and the ethyl acetate layerwas dehydrated with anhydrous sodium sulfate and concentrated todryness. The residue was dissolved in a small quantity of methanol,subjected to a chromatograph tower of Sephadex LH-20 (1.8×15 cm) andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness to give 15.3 mg of the captioned compoundrepresented by the formula (23).

Rf value: 0.22 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent;chloroform-methanol-tetrahydrofuran=5:1:1)

FAB-MS (m/z): 641 (M+H)⁺

¹ H-NMR (200 MHz, DMSO-d₆), δ (ppm): 10.90 (1H, s), 10.38 (1H, s), 9.99(1H, s), 9.30 (1H, s), 8.70 (1H, d, J=8.1 Hz), 8.53 (1H, d, J=8.5 Hz),6.86-7.22 (8H, m), 6.61 (1H, dd, J=2.2, 8.4 Hz), 6.03 (1H, t, J=5.1 Hz),5.43 (1H, d, J=5.4 Hz), 5.35 (1H, t, J=5.0 Hz), 5.22 (1H, d, J=5.4 Hz),4.89 (1H, d, J=5.4 Hz), 4.19 (2H, d, J=5.1 Hz), 4.00 (2H, m), 3.72 (1H,m), 3.53-3.70 (2H, m), 3.38 (1H, m)

EXAMPLE 24

The compound represented by the formula ##STR42##

64.6 mg of the compound obtained in Example A was dissolved in 2 ml ofN,N-dimethylformamide, 30 mg of 2-cyanoethylhydrazine, and the mixturewas stirred at 90° C. for 1.5 hours. 50 ml of 0.2N hydrochloric acid wasadded to the reaction solution, and the mixture was extracted with ethylacetate (50 ml×2). The ethyl acetate layer was concentrated to dryness,and the residue was dissolved in a small quantity of methanol, subjectedto a chromatograph tower of Sephadex LH-20 (1.8×30 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 45.0 mg of the captioned compound represented by theformula (24).

Rf value: 0.39 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=3:1)

FAB-MS (m/z): 588 (M+H)⁺

¹ H-NMR (200 MHz, DMSO-d₆), δ (ppm): 10.91 (1H, s), 10.36 (1H, s), 9.98(1H, s), 8.70 (1H, d, J=8.4 Hz), 8.53 (1H, d, J=8.4 Hz), 7.18 (2H, t,J=8.4 Hz), 6.95-7.10 (3H, m), 6.15 (1H, t, J=4.2 Hz), 5.42 (1H, d, J=5.7Hz), 5.34 (1H, brs), 5.23 (1H, d, J=4.4 Hz), 4.91 (1H, d, J=5.3 Hz),4.00 (2H, m), 3.72 (1H, m), 3.55-3.70 (2H, m), 3.39 (1H, m), 3.30 (2H, td, J=4.2, 6.2 Hz), 2.69 (2H, t, J=6.2 Hz)

EXAMPLE 25

The compound represented by the formula ##STR43##

1.09 g of the compound obtained in Example A was dissolved in 35 ml ofN,N-dimethylformamide--2 ml of water, 455 mg of2-hydrazino-2-imidazoline hydrobromide and 211 mg of sodium bicarbonatewere added, and the mixture was stirred at 80° C. for 2 hours andconcentrated to dryness. The residue was dissolved in 300 ml of 0.2Nhydrochloric acid and extracted with n-butanol (1 L×2). The butanollayer was concentrated to dryness, and the residue was dissolved in asmall quantity of methanol, subjected to a chromatograph tower ofSephadex LH-20 (3.0×80 cm) and eluted with methanol. The fractionscontaining the desired product were concentrated to dryness to give 650mg of the captioned compound represented by the formula (25).

Rf value: 0.55 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; n-butanol:acetic acid:water=4:1:1)

FAB-MS (m/z): 603 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.2 (1H, s), 10.90 (1H, brs),10.50 (1H, s), 10.14 (1H, s), 9.42 (1H, brs), 8.92 (1H, brs), 8.62 (1H,d, J=10.6 Hz), 8.45 (1H, d, J=9.5 Hz), 7.22 (2H, t, J=6.5 Hz), 7.02-7.10(3H, m), 5.48 (1H, d, J=4.7 Hz), 5.32 (2H, brm), 4.94 (1H, d, J=3.5 Hz),4.04 (2H, m), 3.70-3.90 (5H, m), 3.54-3.70 (2H, m), 3.41 (1H, m)

EXAMPLE 26

The compound represented by the formula ##STR44##

48.3 mg of the compound obtained in Example A was dissolved in 1 ml ofN,N-dimethylformamide, 14.3 mg of 1-amino-4-(2-hydroxyethyl) piperazineand 0.1 ml of saturated sodium bicarbonate aqueous solution were added,and the mixture was stirred at 80° C. for 2 hours. This was distributedbetween 50 ml of ethyl acetate and 50 ml of water, 5 ml of 0.2Nhydrochloric acid was added to the water layer, and the mixture wasextracted with n-butanol (100 ml×2). The butanol layer was concentratedto dryness, and the residue was dissolved in a small quantity ofmethanol, subjected to a chromatograph tower of Sephadex LH-20 (1.8×30cm) and eluted with methanol. The fractions containing the desiredproduct were concentrated to dryness to give 22 mg of the captionedcompound represented by the formula (26).

Rf value: 0.53 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; n-butanol:acetic acid:water=4:1:1)

FAB-MS (m/z): 648 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.92 (1H, s), 10.50 (2H, brs),10.10 (1H, s), 8.66 (1H, d, J=7.2 Hz), 8.50 (1H, d, J=8.9 Hz), 7.18 (2H,t, J=8.9 Hz), 7.02-7.12 (3H, m), 5.46 (1H, d, J=5.6 Hz), 5.25-5.40 (3H,brm), 4.86 (1H, d, J=5.6 Hz), 3.95-4.20 (4H, m), 3.70-3.90 (4H, m),3.55-3.70 (4H, m), 3.20-3.50 (6H, m)

EXAMPLE 27

The compound represented by the formula ##STR45##

24 mg of the compound obtained in Example A was dissolved in 0.6 ml ofN,N-dimethylformamide, 10 mg of t-butyl carbazinate acid was added, andthe mixture was stirred at 80° C. for 6 hours. This was mixed with 50 mlof ethyl acetate, the mixture was washed successively with water (30ml×2) and saturated saline, and the ethyl acetate layer was dehydratedwith anhydrous sodium sulfate and concentrated to dryness. The residuewas dissolved in 1 ml of methanol, subjected to a chromatograph tower ofSephadex LH-20 (1.6×20 cm) and eluted with methanol. The fractionscontaining the desired product were concentrated to dryness to give 27.2mg of the captioned compound represented by the formula (27).

Rf value: 0.42 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=4:1)

FAB-MS (m/z): 684 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.99 (1H, s), 10.42 (1H, s), 10.02(1H, s), 9.82 (1H, brs), 8.65 (1H, d, J=7.7 Hz), 8.49 (1H, d, J=7.7 Hz),7.18 (2H, t, J=7.7 Hz), 7.00-7.10 (3H, m), 5.42 (1H, brs), 5.35 (1H,brs), 5.21 (1H, brs), 4.90 (1H, brs), 4.02 (2H, m), 3.72 (1H, m),3.56-3.70 (2H, m), 3.40 (1H, m), 1.50 (9H, s)

EXAMPLE 28

The compound represented by the formula ##STR46## and the compoundrepresented by the formula ##STR47##

177 mg of the compound obtained in Example 1 was dissolved in 6 ml ofN,N-dimethylformamide, 0.68 ml of allyl bromide was added, and themixture was stirred at room temperature for 1 day. 200 ml of water wasadded to the mixture, the mixture was extracted with ethyl acetate (200ml×3), and the ethyl acetate layer was dehydrated with saturated salineand concentrated to dryness. The residue was dissolved in 3 ml ofmethanol, subjected to a chromatograph tower of Sephadex LH-20 (2.5×40cm) and eluted with methanol. The fractions containing the desiredproducts respectively were concentrated to dryness to give 42.1 mg ofthe captioned compound represented by the formula (28) and 67.5 mg ofthe compound represented by the formula (29).

The compound represented by the formula (28)

Rf value: 0.68 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=2:1)

FAB-MS (m/z): 575 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.90 (1H, s), 10.38 (1H, s), 9.98(1H, s), 8.70 (1H, d, J=9.0 Hz), 8.52 (1H, d, J=10.2 Hz), 7.20 (2H, t,J=7.7 Hz), 6.95-7.08 (3H, m), 5.92 (2H, m), 5.40 (1H, d, J=6.4 Hz), 5.32(1H, m), 5.20 (2H, m), 5.05 (1H, d, J=11.5 Hz), 4.88 (1H, d, J=5.8 Hz),4.00 (2H, m), 3.67-3.78 (3H, m), 3.58-3.65 (2H, m), 3.35 (1H, m)

The compound represented by the formula (29)

Rf value: 0.75 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=2:1)

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.91 (1H, s), 10.40 (1H, brs),10.00 (1H, brs), 8.66 (1H, d, J=9.4 Hz), 8.50 (1H, d, J=9.4 Hz), 7.18(2H, t, J=8.0 Hz), 7.00-7.10 (3H, m), 5.90 (2H, ddt, J=6.3, 10.2, 17.0Hz), 5.42 (1H, d, J=5.3 Hz), 5.33 (1H, brs), 5.23 (2H, d, J=17.0 Hz),5.22 (1H, brs), 5.04 (2H, d, J=10.2 Hz), 4.91 (1H, brs), 4.02 (2H, m),3.97 (4H, d, J=6.3 Hz), 3.70 (1H, m), 3.51-3.66 (2H, m), 3.35 (1H, m)

EXAMPLE 29

The compound represented by the formula ##STR48## and the compoundrepresented by the formula ##STR49##

20 mg of the compound obtained in Example 1 was dissolved in 1 ml ofN,N-dimethylformamide, 0.3 ml of benzyl bromide was added, and themixture was stirred overnight. This was mixed with 40 ml of ethylacetate, the mixture was washed successively with water (30 ml×2) andand then saturated saline, and the ethyl acetate layer was dehydratedwith anhydrous sodium sulfate and concentrated to dryness. The residuewas dissolved in 1 ml of methanol, subjected to a chromatograph tower ofSephadex LH-20 (1.6×30 cm) and eluted with methanol. The fractionscontaining the desired products respectively were concentrated todryness to give 13.2 mg of the captioned compound represented by theformula (30) and 7.2 mg of the compound represented by the formula (31).

The compound represented by the formula (30)

Rf value: 0.44 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=3:1)

FAB-MS (m/z): 715 (M+H)⁺

¹ H-NMR (200 MHz, DMSO-d₆), δ (ppm): 10.85 (1H, s), 10.35 (1H, s), 9.96(1H, s), 8.65 (1H, d, J=8.5 Hz), 8.45 (1H, d, J=9.0 Hz), 7.50-7.65 (4H,m), 7.10-7.40 (8H, m), 6.95-7.10 (3H, m), 5.40 (1H, d, J=5.4 Hz), 5.30(1H, brs), 5.18 (1H, d, J=4.9 Hz), 4.83 (1H, d, J=4.9 Hz), 4.58 (2H, s),4.55 (2H, s), 4.00 (2H, m), 3.46-3.80 (3H, m) , 3.36 (1H, m)

The compound represented by the formula (31)

Rf value: 0.38 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=3:1)

FAB-MS (m/z): 625 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.88 (1H, s), 10.40 (1H, brs),10.00 (1H, brs), 8.67 (1H, d, J=7.9 Hz), 8.51 (1H, d, J=7.3 Hz), 7.50(2H, d, J=6.9 Hz), 7.30 (2H, t, J=6.9 Hz), 7.21 (1H, t, J=6.9 Hz), 7.16(2H, t, J=7.3 Hz), 6.96-7.07 (3H, m), 6.13 (1H, t, J=5.3 Hz), 5.42 (1H,d, J=5.9 Hz), 5.21 (1H, d, J=5.3 Hz), 4.91 (1H, brs), 4.55 (1H, brs),4.28 (2H, d, J=5.3 Hz), 4.02 (2H, m), 3.72 (1H, m), 3.55-3.70 (2H, m),3.40 (1H, m)

EXAMPLE 30

The compound represented by the formula ##STR50##

1.4 g of the compound obtained in Example A was dissolved in 30 ml ofN,N-dimethylformamide, 1 g of methyl carbazinate was added, and themixture was stirred at 80° C. for 2 hours. 400 ml of water was added tothe mixture, and the mixture was extracted with ethyl acetate (500ml×3). The resultant ethyl acetate layer was concentrated to dryness.The residue was dissolved in 5 ml of methanol, subjected to achromatograph tower of Sephadex LH-20 (3.0×80 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 1.3 g of the captioned compound represented by theformula (32).

Rf value: 0.18 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=4:1)

FAB-MS (m/z): 592 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.00 (1H, s), 10.42 (1H, brs),10.18 (1H, s), 10.04 (1H, brs), 8.64 (1H, d, J=7.6 Hz), 8.47 (1H, d,J=8.3 Hz), 7.20 (2H, t, J=8.3 Hz), 7.00-7.10 (3H, m), 5.42 (1H, brs),5.35 (1H, brs), 5.21 (1H, brs), 4.91 (1H, brs), 4.02 (2H, m), 3.75 (3H,s), 3.50-3.70 (3H, m), 3.40 (1H, m)

EXAMPLE 31

The compound represented by the formula ##STR51##

90 mg of the compound obtained in Example A was dissolved in 1 ml ofN,N-dimethylformamide, 67 mg of(2R,3S)-3,4-0-isopropylidene-2,3,4-trihydroxybutane carbohydrazide, andthe mixture was stirred at 80° C. for 7 hours and then at roomtemperature for 3 days. 50 ml of water was added to the mixture, and themixture was extracted with ethyl acetate (50 ml×2). The resultant ethylacetate layer was concentrated to dryness. The residue was dissolved in3 ml of methanol, subjected to a chromatograph tower of Sephadex LH-20(1.8×25 cm) and eluted with methanol. The fractions containing thedesired product were concentrated to dryness to give 112 mg of thecaptioned compound represented by the formula (33).

Rf value: 0.14 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=4:1)

FAB-MS (m/z): 692 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.01 (1H, s), 10.70 (1H, s), 10.45(1H, s), 10.05 (1H, s), 8.75 (1H, d, J=7.4 Hz), 8.47 (1H, d, J=7.4 Hz),7.21 (2H, t, J=7.4 Hz), 7.00-7.10 (3H, m), 6.26 (1H, d, J=6.7 Hz), 5.44(1H, d, J=5.9 Hz), 5.39 (1H, brs), 5.24 (1H, d, J=5.9 Hz), 4.93 (1H, d,J=5.9 Hz), 4.31 (1H, dd, J=6.7, 11.9 Hz), 4.22 (1H, t, J=6.7 Hz), 4.10(1H, ddd, J=6.7, 6.7, 11.9 Hz), 4.05 (2H, m), 3.91 (1H, t, J=6.7 Hz),3.76 (1H, m), 3.57-3.71 (2H, m), 3.40 (1H, m), 1.45 (3H, s), 1.36 (3H,s)

EXAMPLE 32

The compound represented by the formula ##STR52##

25 mg of the compound obtained in Example 1 was dissolved in 5 ml ofanhydrous tetrahydrofuran, 10 mg of p-toluenesulfonic anhydride wasadded, and the mixture was stirred at room temperature for 1 day. Thereaction solution was concentrated to dryness, and the residue wasdissolved in 1 ml of methanol, subjected to a chromatograph tower ofSephadex LH-20 (1.8×20 cm) and eluted with methanol. The fractionscontaining the desired product were concentrated to dryness to give 12.3mg of the captioned compound represented by the formula (34).

Rf value: 0.49 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=3:1:1)

FAB-MS (m/z): 688 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.98 (1H, s), 10.87 (1H, s), 10.42(1H, s), 10.05 (1H, s), 8.54 (1H, d, J=7.9 Hz), 8.38 (1H, d, J=7.9 Hz),7.84 (2H, d, J=8.7 Hz), 7.44 (1H, d, J=8.7 Hz), 7.19 (2H, t, J=7.9 Hz),7.00-7.08 (3H, m), 5.43 (1H, d, J=4.7 Hz), 5.35 (1H, brs), 5.23 (1H, d,J=4.9 Hz), 4.90 (1H, d, J=4.4 Hz), 4.04 (2H, m), 3.75 (1H, m), 3.55-3.70(2H, m), 3.40 (1H, m), 2.42 (3H, s)

EXAMPLE 33

The compound represented by the formula ##STR53##

20 mg of the compound obtained in Example 1 was dissolved in 2 ml oftetrahydrofuran, 0.1 ml of phenyl isocyanate was added, and the mixturewas stirred at room temperature for 2 hours. The reaction solution wasconcentrated to dryness, and the residue was dissolved in 1 ml ofmethanol, subjected to a chromatograph tower of Sephadex LH-20 (1.6×30cm) and eluted with methanol. The fractions containing the desiredproduct were concentrated to dryness to give 12 mg of the captionedcompound represented by the formula (35).

Rf value: 0.38 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 653 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.00 (1H, s), 10.40 (1H, brs),10.10 (1H, brs), 9.48 (1H, s), 9.50 (1H, s), 8.67 (1H, d, J=8.3 Hz),8.50 (1H, d, J=8.3 Hz). 7.48 (2H, d, J=7.8 Hz), 7.27 (2H, t, J=7.8 Hz),7.20 (2H, t, J=7.8 Hz), 6.95-7.10 (4H, m), 5.43 (1H, d, J=4.2 Hz), 5.30(1H, brs), 5.23 (1H, brs), 4.95 (1H, brs), 4.03 (2H, m), 3.75 (1H, m),3.58-3.70 (2H, m), 3.38 (1H, m)

EXAMPLE 34

The compound represented by the formula ##STR54##

15 mg of the compound obtained in Example 1 was dissolved in 2 ml oftetrahydrofuran, 16 μl of benzoyl chloride was added, and the mixturewas stirred at room temperature for 2 hours. The solvent was distilledaway, and the residue was dissolved in 1 ml of methanol, subjected to achromatograph tower of Sephadex LH-20 (1.6×20 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 12 mg of the captioned compound represented by theformula (36).

Rf value: 0.57 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 639 (M+H)⁺

¹ H-NMR (200 MHz, DMSO-d₆), δ (ppm): 11.35 (1H, brs), 11.04 (1H, s),10.45 (1H, brs), 10.08 (1H, brs), 8.66 (1H, d, J=8 Hz), 8.49 (1H, d,J=8.5 Hz), 8.04 (2H, d, J=7.1 Hz), 7.55-7.78 (3H, m), 7.20 (2H, t, J=8.5Hz), 7.00-7.15 (3H, m), 5.45 (2H, brs), 5.25 (1H, brs), 4.97 (1H, brs),4.02 (2H, m), 3.55-3.82 (3H, m), 3.41 (1H, m)

EXAMPLE 35

The compound represented by the formula ##STR55##

25 mg of the compound obtained in Example A was dissolved in 1.5 ml ofN,N-dimethylformamide, 30 mg of α-picolinohydrazide was added, and themixture was stirred at 80° C. for 2 hours. This was mixed with 50 ml ofethyl acetate, and the mixture was washed successively with water andthen saturated saline, dehydrated with anhydrous sodium sulfate, andconcentrated to dryness. The residue was dissolved in 1 ml of methanol,subjected to a chromatograph tower of Sephadex LH-20 (1.8×15 cm) andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness to give 30 mg of the captioned compoundrepresented by the formula (37).

Rf value: 0.58 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 640 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.43 (1H, s), 11.02 (1H, s), 10.45(1H, s), 10.07 (1H, s), 8.82 (1H, d, J=4.2 Hz), 8.75 (1H, d, J=7.3 Hz),8.48 (1H, d, J=7.8 Hz), 8.12 (2H, m), 7.75 (1H, m), 7.20 (2H, t, J=7.0Hz), 7.00-7.15 (3H, m), 5.45 (1H, d, J=6.3 Hz), 5.40 (1H, brs), 5.25(1H, d, J=6.3 Hz), 4.96 (1H, brs), 4.04 (2H, m), 3.76 (1H, m), 3.55-3.72(2H, m), 3.42 (1H, m)

EXAMPLE 36

The compound represented by the formula ##STR56##

30 mg of the compound obtained in Example A was dissolved in 1 ml ofN,N-dimethylformamide, 30 mg of 2-hydrazinoethanol was added, and themixture was stirred at 80° C. for 2 hours. This was concentrated todryness. The residue was dissolved in 1 ml of methanol, subjected to achromatograph tower of Sephadex LH-20 (1.8×20 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 32 mg of the captioned compound represented by theformula (38).

Rf value: 0.32 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=2:1)

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.91 (1H, s), 10.35 (1H, brs),9.98 (1H, brs), 8.70 (1H, d, J=6.7 Hz), 8.53 (1H, d, J=6.9 Hz), 7.18(2H, t, J=7.6 Hz), 6.99-7.06 (3H, m), 5.76 (1H, t, J=5.2 Hz), 5.41 (1H,d, J=5.6 Hz), 5.32 (1H, brs), 5.20 (1H, d, J=5.2 Hz), 4.90 (1H, brs),4.51 (1H, t, J=4.9 Hz), 3.96-4.06 (2H, m), 3.73 (1H, m), 3.55-3.70 (4H,m), 3.39 (1H, m), 3.12 (2H, m)

EXAMPLE 37

The compound represented by the formula ##STR57##

40 mg of the compound obtained in Example A was dissolved in 2 ml ofN,N-dimethylformamide, 10 mg of 1-aminopyrrolidine hydrochloride and 0.1ml of sodium bicarbonate aqueous solution were added, and the mixturewas stirred at 80° C. for 2 hours. 40 ml of water was added thereto andthe mixture was extracted with ethyl acetate (40 ml×2). The resultantethyl acetate layer was dehydrated with anhydrous sodium sulfate, andconcentrated to dryness. The residue was dissolved in 1 ml of methanol,subjected to a chromatograph tower of Sephadex LH-20 (1.8×20 cm) andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness to give 10.0 mg of the captioned compoundrepresented by the formula (39).

Rf value: 0.33 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=4:1)

FAB-MS (m/z): 589 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.91 (1H, s), 10.35 (1H, s), 9.95(1H, s), 8.78 (1H, d, J=8.3 Hz), 8.52 (1H, d, J=8.3 Hz), 7.16 (2H, t,J=7.6 Hz), 6.98-7.06 (3H, m), 5.40 (1H, d, J=5.5 Hz), 5.33 (1H, t, J=5.7Hz), 5.18 (1H, d, J=5.5 Hz), 4.85 (1H, d, J=4.8 Hz), 4.02 (2H, m), 3.74(1H, m), 3.53-3.68 (2H, m), 3.30-3.42 (5H, m), 1.97 (4H, m)

EXAMPLE 38

The compound represented by the formula ##STR58##

90 mg of6-benzyloxymethyl-1,11-dibenzyloxy-12,13-dehydro-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-4,7(6H)-dione,a compound disclosed in PCT/WO91/18003, 1.3 g of silver oxide and 550 mgof 4 Å molecular sieve were suspended in 30 ml of anhydrous benzene.After reflux with heating for 20 minutes, a solution of 416.4 mg ofα-bromo-3-deoxy-3-azido-2,4,6-trtacetyl-D-glucose in 5 ml of anhydrousbenzene was added dropwise over a period of 10 minutes. After furtherreflux with heating for 2 days, the insoluble matters were filteredusing Celite. The filtrate was concentrated to dryness, and the residuewas dissolved in 150 ml of ethyl acetate, washed successively with 0.2Nhydrochloric acid, water and then saturated saline, dehydrated withanhydrous sodium sulfate, and concentrated to dryness. The residue wasdissolved in 5 ml of chloroform, subjected to a chromatograph tower ofSephadex LH-20 (3.0×80 cm) and eluted with chloroform. The fractionscontaining the desired product were concentrated to dryness, and theresidue was purified by preparative thin layer chromatography[n-hexane:acetone:tetrahydrofuran=3:1:0.1 (Rf: 0.5), thentoluene:acetone=10:1 (Rf: 0.5)] to give 9.2 mg of6-benzyloxymethyl-1,11-dibenzyloxy-12,13-dehydro-13-(β-D-glucopyranosyl)-5H-indolo[2,3-a]pyrrolo-[3,4-c]carbazole-5,7(6H)-dione.

9.2 mg of the resultant compound was dissolved in 1 ml of hydrazinemonohydrate, and the solution was stirred at room temperature for 4hours. This was mixed with 30 ml of ethyl acetate, and the mixture waswashed successively with 0.2N hydrochloric acid, water and thensaturated saline, dehydrated with anhydrous sodium sulfate, andconcentrated to dryness. The residue was dissolved in 0.5 ml oftetrahydrofuran--1 ml of methanol, palladium black was added, and themixture was stirred, under a hydrogen stream, at room temperature for 3hours. The insoluble matters were filtered using Celite, 1.5 ml of 10%hydrogen chloride-methanol was added to the filtrate, and the mixturewas concentrated to dryness. The residue was dissolved in 0.5 ml ofmethanol, subjected to a chromatograph tower of Sephadex LH-20 (1.0×15cm) and eluted with methanol. The fractions containing the desiredproduct were concentrated to dryness to give 2.0 mg of the captionedcompound represented by the formula (40).

Rf value: 0.5 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; n-butanol:acetic acid:water=4:1:1)

FAB-MS (m/z): 534 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.80 (1H, s), 10.48 (1H, s), 10.20(1H, s), 8.79 (1H, d, J=7.9 Hz), 8.52 (3H, br), 8.50 (1H, d, J=9.2 Hz),7.61 (1H, d, J=6.6 Hz), 7.16 (1H, dd, J=9.2, 9.2 Hz), 7.10 (1H, dd,J=9.2, 9.2 Hz), 7.05 (1H, dd, J=9.2, 9.2 Hz), 7.00 (1H, dd, J=9.2, 9.2Hz), 6.42 (1H, d, J=5.2 Hz), 6.16 (1H, d, J=3.9 Hz), 5.18 (1H, br), 4.93(1H, br), 4.40 (1H, m), 4.16 (1H, m), 4.03 (1H, m), 3.78 (1H, m), 8.68(1H, m), 3.42 (1H, m)

EXAMPLE 39

The compound represented by the formula ##STR59##

30 mg of the compound obtained in Example A was dissolved in 1.5 ml ofN,N-dimethylformamide, 60 mg of cyanoacetohydrazide was added, and themixture was stirred at 80° C. for 9 hours. This was mixed with 30 ml ofethyl acetate, the mixture was washed successively with water and thensaturated saline, and the ethyl acetate layer was dehydrated withanhydrous sodium sulfate and concentrated to dryness. The residue wasdissolved in a small quantity of methanol, subjected to a chromatographtower of Sephadex LH-20 (1.5×15 cm) and eluted with methanol. Thefractions containing the desired product were concentrated to dryness togive 27.8 mg of the captioned compound represented by the formula (41).

Rf value: 0.53 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=3:1:0.1)

FAB-MS (m/z): 601 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.14 (1H, s), 11.01 (1H, s), 10.42(1H, s), 10.04 (1H, s), 8.65 (1H, d, J=7.6 Hz), 8.47 (1H, d, J=7.6 Hz),7.21 (2H, t, J=7.6 Hz), 7.05 (3H, t, J=7.6 Hz), 5.41 (2H, d, J=4.5 Hz),5.19 (1H, d, J=6.8 Hz), 4.90 (1H, d, J=6.8 Hz), 4.13 (2H, s), 4.04 (2H,br), 3.75 (1H, m), 3.64 (2H, m), 3.43 (1H, m)

EXAMPLE 40

The compound represented by the formula ##STR60##

1 g of12,13-dihydro-1,11-dihydroxy-13-(β-D-glucopyranosyl)-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dionewas dissolved in 25 ml of tetrahydrofuran, an ether solution of anexcessive quantity of diazomethane was added, the mixture was stirred at4° C. overnight, and the formed yellow precipitate was collected byfiltration. This was dissolved in 3 ml of hydrazine monohydrate, and thesolution was subjected to reaction at room temperature for 1.5 hours.After the reaction, 200 ml of purified water was added, and theresultant precipitate was collected by filtration, washed successivelywith purified water and then methanol, and dried under reduced pressureto give 683.4 mg of the captioned compound represented by the formula(42).

HPLC; Rt, 10.5 minutes (column: Chromatolex ODS, inner diameter 4.6 mm,length 250 mm, detection; UV 305 nm, flow rate; 1 ml/minute, movingphase; methanol:water=6:4)

FAB-MS (m/z): 563 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, s), 8.87 (1H, d, J=7.8Hz), 8.65 (1H, d, J=7.8 Hz), 7.35 (1H, t, J=7.8 Hz), 7.23 (1H, t, J=7.8Hz), 7.25 (1H, d, J=7.8 Hz), 7.18 (1H, d, J=7.8 Hz), 6.90 (1H, d, J=9.3Hz), 5.40 (1H, brs), 5.18 (1H, brs), 5.00 (2H, brs), 4.90 (2H, brs),4.06 (6H, s), 4.00 (2H, m), 3.78 (1H, m), 3.63 (2H, m), 3.42 (1H, m)

EXAMPLE 41

The compound represented by the formula ##STR61##

708.8 mg of the captioned compound represented by the formula (43) wasobtained from 679 mg of the compound obtained in Example 40, accordingto the process of Example 2.

HPLC; Rt, 10.9 minutes (column: Chromatolex ODS, inner diameter 4.6 mm,length 250 mm, detection; UV 310 nm, flow rate; 1 ml/minute, movingphase; acetonitrile:water=2.8→acetonitrile:water=6:4, 30 minutes lineargradient)

FAB-MS (m/z): 618 [M]⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 13.5 (1H, brs), 11.1 (1H, s), 9.01(1H, s), 8.83 (1H, d, J=7.8 Hz), 8.63 (1H, d, J=7.8 Hz), 7.39 (1H, t,J=7.8 Hz), 7.37 (1H, t, J=7.8 Hz), 7.29 (1H, d, J=7.8 Hz), 7.22 (1.H, d,J=7.8 Hz), 6.94 (1H, d, J=9.3 Hz), 5.43 (1H, d, J=5.4 Hz), 5.22 (1H, d,J=5.4 Hz), 5.01 (1H, brs), 4.93 (1H, d, J=5.4 Hz), 4.07 (6H, s), 4.05(1H, m), 3.96 (1H, m), 3.79 (1H, m), 3.60 (2H, m), 3.44 (1H, m)

EXAMPLE 42

The compound represented by the formula ##STR62##

704 mg of the compound obtained in Example 41 was dissolved in 10 ml ofN,N-dimethylformamide, 60 mg of 10 % palladium-carbon (Pd-C) was added,and the mixture was subjected to hydrogenation at room temperature for 6hours under stirring. The reaction mixture was filtered using a sheet offilter paper on which Celite was spread to remove Pd-C, 200 ml of ethylacetate was added to the filtrate, and the mixture was extracted with 50ml of sodium bicarbonate aqueous solution (pH 8). The water layer wasadjusted to pH 2 and extracted with ethyl acetate (500 ml). The ethylacetate layer was extracted with 2% sodium bicarbonate aqueous solution(70 ml). The 2% sodium bicarbonate aqueous solution layer wasconcentrated under reduced pressure, adsorbed on a column of Dialon HP20 (inner diameter 3 cm, length 30 cm), washed with water, and theneluted with 300 ml of methanol. The methanol eluate was concentrated todryness, the residue was dissolved in a small quantity ofN,N-dimethylformamide, and the solution was subjected to preparativeHPLC (column: Chromatolex ODS, inner diameter 20 mm, length 250 mm,detection; UV 310 nm, flow rate; 9 ml/minute, moving phase;acetonitrile:water=25:75). The fractions containing the desired productwere concentrated to dryness, and the residue was dissolved in a smallquantity of water, subjected to column chromatography of Sephadex G-15(inner diameter 3 cm, length 63 cm) and eluted with water:methanol=9:1.The fractions containing the desired product were concentrated and thenfreeze dried to give 84.2 mg of the sodium salt of the captionedcompound represented by the formula (44).

HPLC; Rt, 8.9 minutes (column: Chromatolex ODS, inner diameter 4.6 mm,length 250 mm, detection; UV 310 nm, flow rate; 1 ml/minute, movingphase; acetonitrile:water:trifluoroacetic acid=300:700:1)

FAB-MS (m/z): 643 (M+Na)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, brs), 8.85 (1H, d, J=7.8Hz), 8.63 (1H, d, J=7.8 Hz), 7.33 (1H, t, J=7.8 Hz), 7.31 (1H, t, J=7.8Hz), 7.24 (1H, d, J=7.8 Hz), 7.16 (1H, d, J=7.8 Hz), 6.89 (1H, d, J=9.3Hz), 5.63 (1H, brs), 5.42 (1H, brs), 5.10 (1H, brs), 4.99 (1H, brs),4.06 (6H, s), 4.02 (2H, m), 3.80 (1H, m), 3.67 (1H, J=8.8 Hz), 3.58 (1H,m), 3.42 (1H, t, J=8.3 Hz), 3.34 (2H, s)

EXAMPLE 43

The compound represented by the formula ##STR63##

23.8 mg of the captioned compound represented by the formula (45) wasobtained from 70 mg of the compound obtained in Example 4, according tothe same process as in Example 2.

FAB-MS (m/z): 641 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.8 (1H, s), 9.26 (1H, d, J=7.8Hz), 9.09 (1H, d, J=7.8 Hz), 8.94 (1H, s), 7.78 (1H, d, J=7.8 Hz), 7.74(1H, d, J=7.8 Hz), 7.50 (2H, t, J=7.8 Hz), 6.98 (1H, d, J=9.3 Hz), 5.44(1H, d, J=5.9 Hz), 5.33 (1H, brs), 5.09 (1H, d, J=5.4 Hz), 3.96 (2H, m),3.85 (1H, m), 3.67 (2H, m), 3.59 (3H, s), 3.56 (1H, m)

EXAMPLE 44

The compound represented by the formula ##STR64##

210 mg of the captioned compound represented by the formula (46) wasobtained from 1 g of the compound obtained in Example 43, according tothe same process as in Example 42.

FAB-MS (m/z): 643 (M+H)⁺

¹ H-NMR (500 MHz, DMSO-d₆), δ (ppm): 10.7 (1H, s), 9.26 (1H, d, J=7.8Hz), 9.09 (1H, d, J=7.8 Hz), 7.74 (1H, d, J=7.8 Hz), 7.71 (1H, d, J=7.8Hz), 7.46 (2H, t, J=7.8 Hz), 6.93 (1H, d, J=9.2 Hz), 6.00 (1H, brs),5.42 (1H, brs), 5.31 (1H, brs), 5.03 (1H, brs), 3.96 (2H, brs), 3.85(2H, s), 3.83 (1H, m), 3.59 (3H, s), 3.50-3.70 (3H, m)

EXAMPLE 45

The compound represented by the formula ##STR65##

48.2 mg of the captioned compound represented by the formula (47) wasobtained from 51.4 mg of the compound obtained in Example 4, accordingto the same process as in Example 5.

FAB-MS (m/z): 613 (M+H)⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 10.9 (1H, brs), 10.8 (1H, brs),9.20 (1H, m), 9.03 (1H, m), 8.48 (1H, s), 7.75 (1H, d, J=7.8 Hz), 7.70(1H, d, J=7.8 Hz), 7.45 (2H, t, J=7.8 Hz), 6.93 (1H, brt, J=9.3 Hz),5.41 (2H, m), 5.04 (1H, d, J=5.9 Hz), 3.99 (2H, brs), 3.86 (1H, m), 3.60(3H, s), 3.52-3.67 (3H, m)

EXAMPLE 46

The compound represented by the formula ##STR66##

13 mg of the captioned compound represented by the formula (48) wasobtained from 14.1 mg of the compound obtained in Example 4, accordingto the same process as in Example 6.

FAB-MS (m/z): 627 (M+H)⁺

¹ H-NMR (500 MHz, DMSO-d₆), δ (ppm): 10.8 (2H, s), 9.20 (1H, m), 9.04(1H, m), 7.74 (2H, m), 7.47 (2H, m), 6.93 (1H, m), 5.41 (1H, m), 5.32(1H, brs), 5.04 (1H, m), 3.96 (2H, brs), 3.85 (1H, m), 3.58 (3H, s),3.50-3.70 (3H, m), 2.12 (3H, s)

EXAMPLE 47

The compound represented by the formula ##STR67##

1 ml of hydrazine monohydrate was added to 3.2 mg of12,13-dihydro-2,10-dihydroxy-13-(β-D-glucopyranosyl)-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione,and the mixture was stirred at room temperature for 2 hours. This wasdistributed with ethyl acetate-0.2N hydrochloric acid, and the ethylacetate layer was washed successively with water then saturated saline,and concentrated to dryness. The residue was dissolved in a smallquantity of methanol, subjected to a chromatograph tower of SephadexLH-20 (1.0×5 cm) and eluted with methanol. The fractions containing thedesired product were concentrated to dryness to give 3.0 mg of thecaptioned compound represented by the formula (49).

Rf value: 0.22 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=3:1:1)

FAB-MS (m/z): 534 [M]⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.16 (1H, s), 9.76 (1H, s), 9.73(1H, s), 8.90 (1H, d, J=7.3 Hz), 8.82 (1H, d, J=7.3 Hz), 7.18 (1H, d,J=2.0 Hz), 6.98 (1H, d, J=2.0 Hz), 6.83 (2H, dt, J=2.0, 7.3 Hz), 5.97(1H, d, J=7.2 Hz), 5.84 (1H, t, J=3.3 Hz), 5.32 (1H, d, J=5.3 Hz), 5.10(1H, d, J=5.3 Hz), 4.93 (1H, d, J=5.2 Hz), 4.90 (2H, s), 4.04-3.86 (2H,m), 3.78 (1H, m), 3.60-3.35 (3H, m)

EXAMPLE 48

The compound represented by the formula ##STR68##

0.4 ml of hydrazine hydrate was added to 7.1 mg of2,10-difluoro-12,13-dihydro-13-(β-D-glucopyranosyl)-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione,and the mixture was stirred at room temperature for 40 minutes. 1.34 mlof concentrated hydrochloric acid was added thereto, and the mixture wasextracted with ethyl acetate. The ethyl acetate layer was washed withwater and concentrated. The residue was dissolved in 3.7 ml ofN,N-dimethylformamide and 0.37 ml of concentrated hydrochloric acid, andthe solution was stirred at room temperature overnight. This wasdistributed between ethyl acetate and water, and the ethyl acetate layerwas concentrated to dryness. The residue was dissolved in a smallquantity of ethanol, subjected to a chromatograph tower of SephadexLH-20 and eluted with ethanol. The fractions containing the desiredproduct were concentrated to dryness to give 4.6 mg of the captionedcompound represented by the formula (50).

FAB-MS (m/z): 566 [M]⁺

¹ H-NMR (400 MHz, DMSO-d₆), δ (ppm): 11.9 (1H, s), 10.8 (1H, brs), 9.07(1H, dd, J=5.8, 8.8 Hz), 9.01 (1H, dd, J=5.9, 8.8 Hz), 8.45 (1H, s),7.93 (1H, brd, J=8.8 Hz), 7.44 (1H, brd, J=8.8 Hz), 7.27 (2H, m), 6. 28(1H, d, J=8.8 Hz), 6.20 (1H, brs), 5.42 (1H, brs), 5.13 (1H, brd, J=5.4Hz), 4.96 (1H, d, J=5.4 Hz), 4.09 (1H, brd, J=7.3 Hz), 3.94 (2H, m),3.83 (1H, brd, J=7.3 Hz), 3.58 (1H, m), 3.45 (1H, m)

EXAMPLE 49

The compound represented by the formula ##STR69## wherein Bn representsa benzyl group.

100 mg of6-benzyloxymethyl-11,11-dibenzyloxy-12,13-dihydro-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione,1.4 g of silver oxide and 0.7 g of 4 Å molecular sieve were suspended in40 ml of anhydrous benzene, the suspension was refluxed with heating for20 minutes, and then a solution of 1-bromo-2,3,5-tri-O-acetyl-D-ribosein 10 ml of anhydrous benzene was added dropwise over a period of 10minutes. The mixture was further refluxed with heating for 3 hours, andthe insoluble matters were filtered using Celite.

The filtrate was concentrated to dryness, and the residue was dissolvedin 100 ml of ethyl acetate and the solution was washed successively with0.2N hydrochloric acid, water and then saturated saline, dried overanhydrous sodium sulfate, and concentrated to dryness. The residue wasdissolved in chloroform, subjected to a chromatograph tower of SephadexLH-20 (2.5×20 cm) and eluted with chloroform. The fractions containingthe desired product were concentrated to dryness, the residue wassubjected to a chromatograph tower of silica gel (2.5×25 cm) and elutedwith toluene-ethyl acetate (3:1), and the fractions containing thedesired product were concentrated to dryness. The residue was furtherpurified by preparative thin layer chromatography (toluene-ethylacetate=5:1 (Rf=0.6)) to give 20.8 mg of6-benzyloxymethyl-1,11-dibenzyloxy-12,13-dihydro-13-(β-D-ribofuranosyl)-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7(6H)-dione.

20.8 mg of this compound was dissolved in 2 ml of hydrazine monohydrate,and the solution was stirred at room temperature for 2 hours. This wasmixed with 30 ml of ethyl acetate, the mixture was washed successivelywith 0.2N hydrochloric acid, water and then saturated saline, andconcentrated to dryness. The residue was dissolved in methanol,subjected to a chromatograph tower of Sephadex LH-20 (1.0×15 cm) andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness, and the residue was purified by preparativethin layer chromatography (chloroform-methanol=10:1 (Rf=0.5)) to give2.9 mg of the captioned compound represented by the formula (51).

Rf value: 0.5 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol=10:1)

FAB-MS (m/z): 684 [M]⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.45 (1H, s), 8.90 (1H, d, J=0.75Hz), 8.68 (1H, d, J=0.75 Hz), 7.18 (2H, d, J=0.75 Hz), 7.11 (2H, d,J=0.75 Hz), 7.20-7.50 (11H, m), 5.35-5.45 (5H, m), 5.17 (1H, d, J=0.38Hz), 5.10 (1H, d, J=0.45 Hz), 4.98 (2H, s), 3.90-4.00 (2H, m), 3.60-3.70(2H, m)

EXAMPLE 50

The compound represented by the formula ##STR70##

33.0 mg of the compound obtained in Example A was dissolved in 3 ml ofN,N-dimethylformamide, 8.4 mg of hydroxyacetohydrazide was added, andthe mixture was stirred at 80° C. for 2 days. This was concentrated todryness, and the residue was dissolved in a small quantity of methanol,subjected to a chromatograph tower of Sephadex LH-20 (1.5×25 cm), andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness, and the residue was dissolved in 30 ml of ethylacetate. The solution was washed with water, and the ethyl acetate layerwas dried over anhydrous sodium sulfate and concentrated to dryness. Theresidue was dissolved in a small quantity of methanol, subjected to achromatograph tower of Sephadex LH-20 (1.5×15 cm) and eluted withmethanol. The fractions containing the desired product were concentratedto dryness to give 29.0 mg of the captioned compound represented by theformula (52).

FAB-MS (m/z): 593 [M+H]⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.00 (1H, s), 10.55 (1H, s), 10.41(1H, s), 10.02 (1H, s), 8.68 (1H, d, J=7.8 Hz), 8.47 (1H, d, J=7.8 Hz),7.20 (2H, t, J=7.8 Hz), 7.04 (3H, m), 5.88 (1H, t, J=7.0 Hz), 5.41 (1H,d, J=6.2 Hz), 5.35 (1H, br), 5.20 (1H, d, J=6.2 Hz), 4.90 (1H, d, J=6.2Hz), 4.16 (2H, d, J=5.7 Hz), 4.03 (2H, m), 3.74 (1H, m), 3.59-3.68 (2H,m), 3.39 (1H, m)

EXAMPLE 51

The compound represented by the formula ##STR71##

35.0 mg of the compound obtained in Example A was dissolved in 1.0 ml ofN,N-dimethylformamide, 35.0 mg of ethylhydrazine oxalate and 0.5 ml ofsaturated sodium bicarbonate aqueous solution were added, and themixture was stirred at 80° C. for 1 day. This was concentrated todryness, and the residue was dissolved in a small quantity of methanol,subjected to a chromatograph tower of Sephadex LH-20 (1.5×15 cm) andeluted with methanol. The fractions containing the desired product wereconcentrated to dryness to give 20.8 mg of the captioned compoundrepresented by the formula (53).

Rf value: 0.5 (produced by Merck Co., Kiesel gel 60F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=2:1:1)

FAB-MS (m/z): 563 [M+H]⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 10.90 (1H, s), 10.35 (1H, s), 9.96(1H, s), 8.72 (1H, d, J=7.9 Hz), 8.54 (1H, d, J=7.9 Hz), 7.17 (2H, t,J=7.9 Hz), 7.03 (3H, m), 5.72 (1H, t, J=4.8 Hz), 5.41 (1H, d, J=6.3 Hz),5.35 (1H, t, J=4.0 Hz), 5.21 (1H, d, J=4.0 Hz), 4.87 (1H, d, J=6.3 Hz),3.96-4.09 (2H, m), 3.73-3.77 (1H, m), 3.58-3.67 (2H, m), 3.37-3.45 (1H,m), 3.07 (2H, m), 1.09 (3H, t, J=7.1 Hz)

EXAMPLE 52

The compound represented by the formula ##STR72##

421.9 mg of the compound obtained in Example C was dissolved in 6.7 mlof N,N-dimethylformamide, 360 mg of 2-hydroxyethylhydrazine was added,and the mixture was stirred at 80° C. for 1.5 hours. The reactionmixture was concentrated to dryness, the residue was dissolved in asmall quantity of methanol, and the solution was placed on a SephadexLH-20 (6.5×50 cm) column for chromatography and eluted with methanol.Fractions containing the desired product were concentrated to dryness togive 189.3 mg of the captioned compound represented by the formula (54).

Rf value: 0.50 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; chloroform:methanol:tetrahydrofuran=3:1:1) FAB-MS (m/z) : 579(M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 11.20 (1H, s), 9.88 (1H, s), 9.85(1H, s), 8.85 (1H, d, J=9.0 Hz), 8.79 (1H, d, J=9.0 Hz), 7.19 (1H, d,J=1.4 Hz), 6.98 (1H, d, J=1.4Hz), 6.82 (1H, dd, J=1.4 Hz, 9.0 Hz), 6.79(1H, dd, J=1.4 Hz, 9.0 Hz), 5.98 (1H, d, J=9.0 Hz), 5.85 (1H, t, J=4.0Hz), 5.73 (1H, t, J=3.4 Hz), 5.31 (1H, d, J=3.6 Hz), 5.10 (1H, d, J=6.3Hz), 4.90 (1H, t, J=4.5 Hz), 3.87-4.10 (3H, m), 3.77-3.80 (1H, m),3.49-3.60 (4H, m), 3.05-3.15 (2H, m)

EXAMPLE 53

The compound represented by the formula ##STR73##

1) 6.73 g of DL-glyceraldehyde dimer and 9.87 g of carbazi acidtert-butyl ester were dissolved in 50 ml of 95% ethanol, and thesolution was stirred at room temperature for 15 hours and then at 60° C.for 30 minutes. The reaction solution was concentrated under reducedpressure, and the residue was recrystallized from ethyl acetate to give13.7 g of (2SR)-3-(tert-butyloxycarbonyl)hydrazono-1,2-propanediol as acolorless solid.

Rf value: 0.49 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; dichloromethane:methanol=10:2)

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 1.41 (9H, s), 3.39 (2H, m), 3.93(1H, m), 4.65 (1H, t, J=5.8 Hz), 5.11 (1H, d, J=5.0 Hz), 7.16 (1H, d,J=6.0 Hz), 10.52 (1H, brs)

2) 52 ml of borane-tetrahydrofuran complex was added to 12.6 g of(2SR)-3-(tert-butyloxycarbonyl)hydrazono-1,2-propanediol at roomtemperature, and the mixture was stirred at room temperature for 10minutes. 26 ml of 6N hydrochloric acid was added to the reactionsolution at room temperature, and the mixture was stirred at 100° C. for15 minutes. The reaction solution was concentrated under reducedpressure, and the residue was adsorbed on an H⁺ type of Dowex 50 WX4,washed with water and eluted with 0.5N ammonia water. Fractionscontaining the desired product were combined and concentrated underreduced pressure, and the resultant oily matter was adsorbed on an NH₄ ⁺type of IRC-50 and eluted with water. Fractions containing the desiredproduct were combined and concentrated under reduced pressure to give2.2 g of (2SR)-3-hydrazino-1,2-propanediol as a colorless solid.

Rf value: 0.33 (producing by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; n-butanol:acetic acid:water=5:2:1)

FAB-MS (m/z): 107 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 2.51 (1H, dd, J=7.3, 12.1 Hz), 2.66(1H, dd, J=4.3, 12.1 Hz), 3.28 (2H, m), 3.56 (1H, m)

3) 500 mg of the compound obtained in Example C and 313 mg of(2SR)-3-hydrazino-1,2-propanediol obtained in 2) were dissolved in 10 mlof N,N-dimethylformamide, and the solution was shirred at 80° C. for 2.5hours. The reaction solution was concentrated under reduced pressure,and the residue was purified by Sephadex LH20(chloroform:methanol:ethanol:water=5:2:2:1) to give 462 mg of thecaptioned compound represented by the formula (55).

Rf value: 0.31 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; tetrahydrofuran:methanol:n-hexane:formic acid=10:2:4:0.1)

FAB-MS (m/z): 609 (M+H)⁺

[α]_(D) +172° (c=1.02, DMSO)

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 2.99 (1H, m), 3.07 (1H, m), 3.42(2H, t, J=5.6 Hz), 3.50 (2H, m), 3.62 (1H, m), 3.78 (1H, m), 3.91 (2H),4.01 (1H, m), 4.52 (1H, t, J=5.6 Hz), 4.60 (1H, d, J=4.4 Hz), 4.92 (1H,d, J=5.2 Hz), 5.12 (1H, d, J=5.2 Hz), 5.33 (1H, d, J=4.4 Hz), 5.71 (1H,dd, J=4.4 Hz, 5.6 Hz), 5.87 (1H, m), 5.97 (1H, d, J=8.5 Hz), 6.80 (1H,dd, J=2.0 Hz, 8.6 Hz), 6.83 (1H, dd, J=1.9 Hz, 8.6 Hz), 6.98 (1H, d,J=2.0 Hz), 7.18 (1H, d, J=1.9 Hz), 8.79 (1H, d, J=8. 6 Hz), 8.87 (1H, d,J=8.6 Hz), 9.76 (1H, s), 9.78 (1H, s), 11.20 (1H, s)

EXAMPLE 54

The compound represented by the formula ##STR74##

1) 3.45 g of D-(R)-glyceraldehyde acetonide and 3.50 g carbazinic acidtert-butyl ester were dissolved in 20 ml of ethanol, and the solutionwas stirred at room temperature for 15 hours. The reaction solution wasconcentrated under reduced pressure, and the residue was recrystallizedfrom ethyl acetate to give 5.65 g of (4S)-2,2-dimethyl-4-(tert-butyloxycarbonylhydrazono)methyl-1,3-dioxolane asa colorless solid.

Rf value: 0.56 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; n-hexane:ethyl acetate=1:1)

[α]_(D) +46.4° (c=1.00, CHCl₃)

¹ H-NMR (300 MHz, CDCl₃), δ (ppm): 1.38 (3H, s), 1.43 (3H, s), 1.50 (9H,s), 3.87 (1H, dd, J=6.3 Hz, 8.6 Hz), 4.18 (1H, dd, J=6.6 Hz, 8.6 Hz),4.65 (1H, ddd, J=6.4 Hz, 6.4 Hz, 6.4 Hz), 7.12 (1H, brd, J=5.9 Hz), 7.78(1H, brs)

2) 18.4 ml of borane-tetrahydrofuran complex was added to 3.00 g of(4S)-2,2-dimethyl-4-(tert-butyloxycarbonylhydrazono)methyl-1,3-dioxolaneat room temperature, and the mixture was stirred at room temperature for10 minutes. 6.2 ml of 6N hydrochloric acid was added to the reactionsolution at room temperature, and mixture was stirred at 100° C. for 15minutes. The reaction solution was concentrated under reduced pressure,and the resultant residue was adsorbed on an H⁺ type of Dowex 50 WX4,washed with water and eluted with 0.5N ammonia water. Fractionscontaining the desired product were combined and concentrated underreduced pressure, and the resultant oily matter was adsorbed on an NH₄ ⁺type of IRC-50 and eluted with water. Fractions containing the desiredproduct were combined, and concentrated under reduced pressure to give772 mg of (2S)-3-hydrazino-1,2-propanediol as a colorless solid.

Rf value: 0.33 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; n-butanol:acetic acid:water=5:2:1)

[α]_(D) -23.0° (c=1.00, MeOH)

¹ H-NMR (300 MHz, CD₃ OD), δ (ppm): 2.72 (1H, dd, J=8.2 Hz, 12.3 Hz),2.87 (1H, dd, J=3.7 Hz, 12.3 Hz), 3.49 (2H, d, J=5.4 Hz), 3.77 (1H, m)

3) 500 mg of the compound obtained in Example C and 306 mg or(2S)-3-hydrazino-1,2-propanediol obtained in 2) were dissolved in 20 mlof N,N-dimethylformamide, and the solution was stirred at 80° C. for 3hours. The reaction solution was concentrated under reduced pressure,and residue was purified by Sephadex LH20(chloroform:methanol:ethanol:water=5:2:2:1) to give 340 mg or thecaptioned compound represented by the formula (56).

Rf value: 0.59 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; tetrahydrofuran:methanol:n-hexane:formic acid=10:2:2:0.1)

[α]_(D) +147° (c=1.00, DMSO)

FAB-MS; (M+H)⁺ ; Found: 609, 1848, Calcd: 609, 1833

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 2.98 (1H, m), 3.07 (1 H, m), 3.41(2H, brt, J=5.7 Hz), 3.50 (2H, m), 3.63 (1H, m), 3.78 (1H, m), 3.85-3.96(2H), 4.02 (1H, m), 4.52 (1H, t, J=5.7 Hz), 4.61 (1H, d, J=4.4 Hz), 4.93(1H, d, J=5.1 Hz), 5.12 (1H, d, J=5.0 Hz), 5.34 (1H, d, J=4.8 Hz), 5.71(1H, m), 5.87 (1H, t, J=4.5 Hz), 5.98 (1H, d, J=8.5 Hz), 6.80 (1H, dd,J=2.2 Hz, 8.7 Hz), 6.83 (1H, dd, J=2.0 Hz, 8.5 Hz), 6.98 (1H, d, J=2.0Hz), 7.18 (1H, d, J=1.9 Hz), 8.78 (1H, d, J=8.7 Hz), 8.87 (1H, d, J=8.6Hz), 9.76 (1H, s), 9.79 (1H, s), 11.20 (1H, s)

EXAMPLE 55

The compound represented by the formula ##STR75##

1) 8.5 g of (S)-(-)-glycidol was dissolved in 80 ml of ethanol, 45.5 gof carbazinic acid tert-butyl ester was added, and the mixture wasstirred at room temperature for 3 days. The reaction solution wasconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (dichloromethane:methanol=10:1) to give5.53 g of (2R)-3-(N'-tert-butyloxycarbonyl)hydrazino-1,2-propanediol aslight yellow oily matter.

Rf value: 0.24 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; dichloromethane:methanol=10:1)

[α]_(D) +10.8° (c=1.00, MeOH)

¹ H-NMR (300 MHz, CD₃ OD), δ (ppm): 1.44 (9H, s), 2.75 (1H, dd, J=7.9Hz, 12.3 Hz), 2.87 (1H, dd, J=4.2 Hz, 12.3 Hz), 3.51 (2H, m), 3.69 (1H,m)

2) 2.99 g of (2R)-3-(N'-tert-butyloxycarbonyl)hydrazino-1,2-propanediolwas dissolved in 15 ml of tetrahydrofuran, 7.5 ml of 6N hydrochloricacid was added, and the mixture was stirred at 100° C. for 20 minutes.The reaction solution was concentrated under reduced pressure, and theresultant residue was adsorbed on an H⁺ type of Dowex 50 WX4, washedwith water and then eluted with 0.5N ammonia water. Fractions containingthe desired product were combined and concentrated under reducedpressure, and the resultant oily matter was adsorbed on an NH₄ ⁺ type ofIRC-50 and eluted with water. Fractions containing the desired productwere combined and concentrated under reduced pressure to give 993 mg of(2R)-3-hydrazino-1,2-propanediol as a colorless solid.

Rf value: 0.33 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; n-butanol:acetic acid:water=5:2:1)

[α]_(D) +20.6° (c=0.992, MeOH)

¹ H-NMR (300 MHz, CD₃ OD), δ (ppm): 2.72 (1H, dd, J=8.2, 12.4 Hz), 2.87(1H, dd, J=3.7, 12.4 Hz), 3.49 (2H, d, J =5.5 Hz), 3.77 (1H, m)

3) 250 mg of the compound obtained in Example C and 129 mg of(2R)-3-hydrazino-1,2-propanediol obtained in 2) were dissolved in 7.5 mlof N,N-dimethylformamide, and the solution was stirred at 80° C. for 3hours. The reaction solution was concentrated under reduced pressure,and the residue was purified by Sephadex LH20(chloroform:methanol:ethanol:water=5:2:2:1) to give 172 mg of thecaptioned compound represented by the formula (57).

Rf value: 0.57 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; tetrahydrofuran:methanol:n-hexane:formic acid=10:2:2:0.1)

[α]_(D) +170° (c=0.50, DMSO)

FAB-MS; (M+H)⁺ ; Found: 609, 1836, Calcd: 609, 1833

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 2.98 (1H, m), 3.07 (1H, m), 3.42(2H, br), 3.51 (2H, br), 3.62 (1H, br), 3.78 (1H, brd, J=10 Hz),3.85-3.96 (2H), 4.02 (1H, m), 4.52 (1H, br), 4.60 (1H, br), 4.93 (1H,br), 5.13 (1H, br), 5.38 (1H, br), 5.72 (1H, t, J=4.8 Hz), 5.90 (1H,br), 5.97 (1H, d, J=8.2 Hz), 6.80 (1H, dd, J=2.1, 8.6 Hz), 6.83 (1H, dd,J=1.9 Hz, 8.6 Hz), 6.99 (1H, d, J=2.0 Hz), 7.18 (1H, d, J=1.8 Hz), 8.79(1H, d, J=8.6 Hz), 8.87 (1H, d, J=8.6 Hz), 9.8 (2H, br), 11.20 (1H, s)

EXAMPLE 56

The compound represented by the formula ##STR76##

1) 10.0 of dihydroxyacetone dimer and 14.7 g of carbazi acid tert-butylester were dissolved in 500 ml of ethanol, and the solution was stirredat room temperature for 15 hours. The reaction solution was concentratedunder reduced pressure, and the residue was recrystallized from ethylacetate to give 18.67 g of2-(tert-butyloxycarbonyl)hydrazono-1,3-propanediol as a colorless solid.

Rf value: 0.49 (produced by Merck Co., Kiesel gel 60 F₂₅₄, developingsolvent; dichloromethane:methanol=10:1.5)

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 1.49 (9H, s), 3.92 (2H, d, J=5.2Hz), 4.24 (2H, d, J=5.0 Hz), 4.88 (1H, t, J=5.8 Hz), 5.61 (1H, t, J=5.1Hz), 9.98 (1H, brs)

2) 50 ml of borane-tetrahydrofuran complex was added to 5.00 g of2-(tert-butyloxycarbonyl)hydrazono-1,3-propanediol at 0° C. and themixture was stirred at room temperature for 0.5 hour. 25 ml of 6Nhydrochloric acid was added to the reaction solution at roomtemperature, and the mixture was refluxed with heating for 1.5 hours.The reaction solution was concentrated under reduced pressure, and theresultant residue was adsorbed on an H⁺ type of Dowex 50 WX4, washed andeluted with 0.5N ammonia water. Fractions containing the desired productwere combined and concentrated under reduced pressure, and the resultantoily matter was adsorbed on an NH₄ ⁺ type of IRC-50 and eluted withwater. Fractions containing the desired product were combined andconcentrated under reduced pressure to give 2.26 g of2-hydrazino-1,3-propanediol as a colorless solid.

FAB-MS (m/z): 107 (M+H)⁺

¹ H-NMR (200 MHz, CD₃ OD), δ (ppm): 2.78 (1H, m), 3.50-3.75 (4H, m)

3) 500 mg of the compound obtained in Example C and 152 mg of2-hydrazino-1,3-propanediol obtained in 2) were dissolved in 50 ml ofN,N-dimethylformamide, and the solution was stirred at 80° C. for 1hour. The reaction solution was concentrated under reduced pressure, andthe residue was purified by Sephadex LH20(chloroform:methanol:ethanol:water=5:2:2:1) to give 418 mg of thecaptioned compound represented by the formula (58).

Rf value: 0.20 (produced by Merck Co,. Kiesel gel 60 F₂₅₄, developingsolvent; dichloromethane:methanol:tetrahydrofuran=3:1:1)

FAB-MS (m/z): 609 (M+H)⁺

¹ H-NMR (300 MHz, DMSO-d₆), δ (ppm): 3.2-3.3 (1H, m), 3.4-3.6 (6H, m),3.78 (1H, m), 3.85-3.95 (2H), 4.02 (1H, m), 4.53 (2H, t, J=5.4 Hz), 4.91(1H, d, J=5.1 Hz), 5.11 (1H, d, J=5.3 Hz), 5.32 (1H, d, J=4.6 Hz), 5.55(1H, d, J=2.6 Hz), 5.86 (1H, t, J=3.8 Hz), 5.97 (1H, d, J=8.3 Hz), 6.80(1H, dd, J=2.0 Hz, 8.6 Hz), 6.82 (1H, dd, J=2.0 Hz, 8.6 Hz), 6.98 (1H,d, J=2.0 Hz), 7.18 (1H, d, J=1.7 Hz), 8.79 (1H, d, J=8.6 Hz), 8.87 (1H,d, J=8.6 Hz), 9.75 (1H, s), 9.78 (1H, s), 11.20 (1H, s).

EXAMPLE 57

Excellent antitumor activities of the compounds (54) to (58) obtained inExamples 52 to 56 are exhibited by the following pharmacological tests.

(1) Proliferation inhibition activity (CTX) against various cancer cellsin culture

Measurement method:

100 μportions of a cell culturing medium (10% fetal bovineserum-containing-RPMI-1640 medium) containing 3×10³ mouse leukemia cell(P388), human gastric cancer cell (MKN-45), human lung cancer cell(PC-13) or human colon cancer cell (DLD-1) were put in a 96-wellmicroplate, the cells were cultured under 5% CO₂ at 37° C. for 24 hours,10 μl each of test solutions containing test compounds respectively wereadded respectively, and the cells were further cultured under 5% CO₂ at37° C. for 72 hours. 10 μl portions of 0.5% Thiazoyl Blue were added tothe culture medium, and incubation was carried out under 5% CO₂ at 37°C. for 2 hours to carry out enzymatic reaction. 20% sodium dodecylsulfate (SDS) was added to discontinue the reaction, incubation wasfurther carried out at 37° C. for 4 hours to dissolve the formed dye,and absorbance at 550 nm was measured and compared with time controlgroup. The results are shown in the following Table 4.

                  TABLE 4                                                         ______________________________________                                        Proliferation inhibition activities                                           (CTX) against various cancer cells                                            Test               CTX (μM)                                                compound   P388    MKN-45     PC-13 DLD-1                                     ______________________________________                                        Example 52 0.00048 0.0017     0.0058                                                                              0.058                                     Example 53 0.0022  0.0069     0.014 0.025                                     Example 54 0.0014  0.0066     0.037 0.039                                     Example 55 0.0012  0.0050     0.019 0.057                                     Example 56 0.0015  0.0048     0.056 0.12                                      ______________________________________                                    

(2) Growth inhibitory effects against human gastric cancer MKN-45 inmice

MKN-45 solid tumor previously subcutaneously implanted into a nude mouseand grown was cut into pieces, and the pieces each 3 mm cubic weresubcutaneously implanted into test mice, respectively. From the timewhen each tumor grew to 0.3 cm³ after the implantation, predeterminedquantities of test chemicals were injected into the tail veins of themice once a day for continuous five days, and after recuperation for 2days, further injected for 5 days (treatment schedule: 5/w×2) or fourtimes every 3 to 4 days (treatment schedule: 2/w×2) for treatment,respectively. 20 days or 32 days after the start of treatment, eachtumor was taken out, and weighed whereby tumor growth inhibition ratioswere calculated and total doses (GID₇₅, mg/kg) to inhibit tumor growthto an extent of 75% were calculated. The results are shown in thefollowing Table 5.

                  TABLE 5                                                         ______________________________________                                        Growth inhibitory effects of compounds of                                     this invention on human gastric cancer                                        MKH-45 in mice                                                                Test           Treatment                                                                              GID.sub.75                                            compound       schedule (mg/kg total)                                         ______________________________________                                        Example 52     5/w × 2                                                                          12                                                    Example 52     2/w × 2                                                                          7.2                                                   Example 56     2/w × 2                                                                          3.0                                                   ______________________________________                                    

EXAMPLE 58

50 g of the compound of Example 5 was dissolved in a solution wherein600 g of macrogol 400 of the Japanese Pharmacopoeia was dissolved in 400g of distilled water for injection, and the solution was filtered forremoval of bacteria using a filter of 0.2 μm. 5 ml portions of thefiltrate were filled into washed and sterilized vials according to aconventional method, and the vials were stopped and capped to give aninjection containing 250 mg of the compound of Example 5 per vial.Administration is made using an agent for intravenous drip wherein 5 to10 ml of this injection (250 to 500 mg of the compound of Example 5) wasadded to and diluted with 500 ml of an infusion such as 5% glucose.

The compounds of formula (I) according to this invention not only haveexcellent anti-tumor activity as evidenced by the foregoing data, butalso are highly advantageous over rebeccamycin and other knownrebeccamycin derivatives, such as BE-13793C, in view of their greatlyenhanced water solubility. This water solubility allows the inventioncompounds to be dissolved in distilled water and other aqueous solventsuseful in formulating, for instance, intravenous injection compositions.The enhanced water solubility may be appreciated from the followingwater solubility test.

EXAMPLE 59

I. Water solubility test:

Water solubility was compared between the compounds in Examples 2, 3, 5,6, 8-11, 13, 18, 20, 25, 30, 38 and 41-44 of the present specification,and the rebeccamycin disclosed in Richard P. Joyce et al., J. Org.Chem., Vol.52, No.7, 1987, pp. 1177-1185, page 1178, as Compound 3 whichis represented by the following formula: ##STR77##

I-1. Test method:

Test samples (200 μg) were suspended in 1.0 ml of water and shaked for10 min. and then centrifuged at 3,000 rpm for 10 min. When precipitateswere observed, supernatant fraction was quantitated by HPLC method.

Column; Chromatorex ODS (4.6×250 mm)

Mobile Phase; 80% MeOH

Flow rate; 1.0 ml/min

Detection; UV 320 nm

I-2. Results:

The results are shown in Table A.

                  TABLE A                                                         ______________________________________                                        Water solubility of rebeccamycin and                                          Example compounds.                                                            Compound         Water solubility (mg/ml)                                     ______________________________________                                        rebeccamycin     0.001                                                        Compound of Example 2                                                                          >0.2                                                         Compound of Example 3                                                                          >0.2                                                         Compound of Example 5                                                                          0.2                                                          Compound of Example 6                                                                          >0.2                                                         Compound of Example 8                                                                          >0.2                                                         Compound of Example 9                                                                          >0.2                                                         Compound of Example 10                                                                         >0.2                                                         Compound of Example 11                                                                         >0.2                                                         Compound of Example 13                                                                         >0.2                                                         Compound of Example 18                                                                         >0.2                                                         Compound of Example 20                                                                         >0.2                                                         Compound of Example 25                                                                         >0.2                                                         Compound of Example 30                                                                         >0.2                                                         Compound of Example 38                                                                         >0.2                                                         Compound of Example 41                                                                         >0.2                                                         Compound of Example 42                                                                         >0.2                                                         Compound of Example 43                                                                         >0.2                                                         Compound of Example 44                                                                         >0.2                                                         ______________________________________                                    

The compounds of this invention also have improved antitumor activity ascompared to other known water soluble derivatives of rebeccamycin asshown in the following example.

EXAMPLE 60

Antitumor activity was compared between the compounds shown in Examples1, 2, 5, 6, 9, 11, 24, 25, 30, 36, 39 and 50 and 6-(2-diethylaminoethyl)rebeccamycin (hereinafter called "DEAE-rebeccamycin") mentioned in TheJournal of Antibiotics, Vol.43, No.1, pp 125-127, which is representedby the following formula: ##STR78##

Test method:

Therapeutic effect of a test compound against mouse tumor (p 388) wasdetermined in the same manner as in the present specification, page 20,line 22 to page 21, line 13.

The tests were conducted four times, and a group to which no medicinewas administered was employed as a control.

The results of the tests are shown in the following Tables B-E:

                  TABLE B                                                         ______________________________________                                        Dose      DEAE-rebeccamycin                                                                           Compound of Example 5                                 mg/kg/day MST.sup.1)  (T/C %).sup.2)                                                                  MST (T/C %)                                           ______________________________________                                        0         10.6 (100)    12.3 (100)                                            0.3       --            16.8 (137)                                            0.63      11.6 (109)    --                                                    1.0       --            16.8 (137)                                            2.5       13.2 (125)    --                                                    3.0       --            17.8 (145)                                            10        14.8 (140)    26.6 (216)                                            30        --            >27.4 (>223)                                          40        13.4 (126)    --                                                    100       toxic         >52.4 (>426)                                          ______________________________________                                    

                  TABLE C                                                         ______________________________________                                                       Dose        MST                                                Compound       mg/kg       day    T/C %                                       ______________________________________                                        Control        --          12.8   100                                         Compound of Example 1                                                                        10          18.2   142                                                        30          >31.0  >242                                                       100         >52.8  >413                                        Compound of Example 2                                                                        1           15.8   128                                                        3           17.8   145                                                        10          >26.4  >245                                                       30          >42.2  >343                                                       100         >47.2  >384                                        ______________________________________                                    

                  TABLE D                                                         ______________________________________                                                        Dose       MST                                                Compound        mg/kg      day    T/C %                                       ______________________________________                                        Control         --         13.0   100                                         Compound of Example 5                                                                         1          21.2   163                                                         3          30.4   234                                                         10         26.5   204                                         Compound of Example 6                                                                         3          24.8   191                                                         10         >28.8  >222                                                        30         >44.0  >338                                                        100        >43.0  >331                                        Compound of Example 9                                                                         1          25.4   195                                                         3          >32.6  >251                                                        10         >60.0  >462                                        Compound of Example 11                                                                        3          21.4   165                                                         10         24.0   185                                                         30         >32.8  >252                                        Compound of Example 25                                                                        1          >29.8  >229                                                        3          >34.8  >268                                                        10         >54.2  >417                                                        30         >56.6  >435                                        Compound of Example 30                                                                        3          22.0   169                                                         10         >27.3  >210                                                        30         >43.2  >332                                                        100        >52.6  >405                                        ______________________________________                                    

                  TABLE E                                                         ______________________________________                                                        Dose       MST                                                Compound        mg/kg      day    T/C %                                       ______________________________________                                        Control         --         11.0   100                                         Compound of Example 5                                                                         12.5       19.0   172                                                         50         25.0   227                                         Compound of Example 24                                                                        3.13       15.0   136                                                         12.5       17.0   154                                                         50         26.0   236                                         Compound of Example 36                                                                        3.13       15.0   136                                                         12.5       20.0   181                                                         50         28.0   254                                         Compound of Example 39                                                                        12.5       17.0   154                                                         50         26.0   236                                         Compound of Example 50                                                                        12.5       18.0   163                                                         50         27.0   245                                         ______________________________________                                    

1) MST=Mean Survival Time in Days

2) T/C=(MST Treated/MST Control)×100%

CDF1 mice were inoculated intraperitoneally (i.p.) with 10⁸ P388 cellson day 0 and treated with each sample consecutively from day 1 for 10days (Q1D for day 1-10). Ten mice were used for control group and 5 micewere used for each dose.

We claim:
 1. A process for the preparation of a compound of the formula(I) ##STR79## wherein R¹ and R² each, independently, represent ahydrogen atom, C₁ -C₈ alkyl group, C₃ -C₆ alkenyl group, C₆ -C₁₂aromatic hydrocarbon group, C₁ -C₆ alkyl group substituted by C₆ -C₁₂aromatic hydrocarbon group or 5- or 6-membered nonaromatic or aromaticheterocyclic group containing 1 to 4 hetero atoms selected from thegroup consisting of nitrogen atom, oxygen atom and sulfur atom, whereinthe C₁ -C₆ alkyl group, C₃ -C₆ alkenyl group, C₃ -C₆ alkynyl group, C₆-C₁₂ aromatic hydrocarbon group, C₁ -C₆ alkyl group substituted by C₆-C₁₂ aromatic hydrocarbon group and heterocyclic group may besubstituted by 1 to 5 substituents selected from the group consisting ofcarboxyl group, carbamoyl group, SO₃ H, NH₂, cyano group, mono-(C₁ -C₆alkyl) amino group, di(C₁ -C₆ alkyl)amino group, hydroxyl group, andhalogen atom; or represent a group of the formula --Y--R³, wherein Yrepresents a carbonyl group, thiocarbonyl group or sulfonyl group, andR³ represents a hydrogen atom, C₁ -C₆ alkyl group, C₃ -C₆ cycloalkylgroup, C₄ -C₁₀ cycloalkylalkyl group, C₆ -C₁₂ aromatic hydrocarbongroup, C₁ -C₆ alkyl group substituted by C₆ -C₁₂ aromatic hydrocarbongroup, C₁ -C₆ alkoxy group, NHNH₂, NH₂, amino group substituted by C₆-C₁₂ aromatic hydrocarbon group, carbamoyl group or 5- or 6-memberednonaromatic or aromatic heterocyclic group containing 1 to 4 heteroatoms selected form the group consisting of nitrogen atom, oxygen atomand sulfur atom, wherein the C₁ -C₆ alkyl group, C₃ -C₆ cycloalkylgroup, C₄ -C₁₀ cycloalkylalkyl group, C₆ -C₁₂ aromatic hydrocarbongroup, C₁ -C₆ alkyl group substituted by C₆ -C₁₂ aromatic hydrocarbongroup and 5- or 6-membered nonaromatic or aromatic heterocyclic groupmay be substituted by 1 to 4 substituents selected from the groupconsisting of halogen atom, unprotected hydroxyl group, hydroxyl groupprotected by a protective group selected from the group consisting of C₂-C₆ alkanoyl group, C₂ -C₇ alkoxycarbonyl group, benzoyl group,acetal-forming group, benzyl group and benzyl group substituted by C₁-C₆ alkoxy group, nitro group, NH₂, carboxyl group, carbamoyl group,cyano group or C₂ -C₇ alkoxycarbonyl group, and the NH₂ and carbamoylgroup are unsubstituted or mono- or di-substituted by unsubstituted C₁-C₆ alkyl group(s) or by C₁ -C₆ alkyl group(s) substituted bysubstituent(s) selected from the group consisting of halogen atom,hydroxyl group, NH₂, carboxyl group, carbamoyl group and C₂ -C₇alkoxycarbamoyl group; orR¹ and R² combine to form an unsubstituted C₁-C₆ alkylidene group or a C₁ -C₆ alkylidene group substituted by 1 to 4substituents selected form the group consisting of NH₂, mono-(C₁ -C₆alkyl)amino group, di-(C₁ -C₆ alkyl)amino group, hydroxyl group,carboxyl group and SO₃ H; or R¹ and R² combine together with thenitrogen atom to which they are bonded to form a 5- or 6-memberednonaromatic heterocyclic group containing 1 to 4 hetero atoms selectedfrom the group consisting of nitrogen atom, oxygen atom and sulfur atom,and the heterocyclic group may be substituted on the ring byunsubstituted C₁ -C₆ alkyl group(s) and by C₁ -C₆ alkyl groupssubstituted by group(s) selected form the group consisting of NH₂,hydroxyl group, carboxyl group and SO₃ H; G represents pentose group orhexose group, and X¹ and X² each, independently, represent a hydrogenatom, halogen atom, NH₂, mono-(C₁ -C₆ alkyl)amino group, di-(C₁ -C₆alkyl)amino group, hydroxyl group, C₁ -C₆ alkoxy group, C₁ -C₆ alkoxygroup substituted by C₆ C₁₂ aromatic hydrocarbon group, carboxyl group,C₂ -C₇ alkoxycarbonyl group or C₁ -C₆ alkyl group, or a pharmaceuticallyacceptable salt thereof, which comprises reacting a compound representedby the following formula (II) or formula (III) wherein any functionalgroup may be protected, ##STR80## wherein Z represents a hydrogen atomor substituted or unsubstituted lower alkyl group, and X¹, X² and G havethe same meaning as defined above,with a compound represented by thefollowing formula (IV) or a derivative thereof wherein functional groupsare protected, ##STR81## wherein R¹³ and R²³ each independentlyrepresent a hydrogen atom, lower alkyl group, lower alkenyl group, loweralkynyl group, aryl group, aralkyl group or heterocyclic group, whereinthe lower alkyl group, lower alkenyl group, lower alkynyl group, arylgroup, alkyl group and heterocyclic group may have 1 to 5 substitutentsselected from the group consisting of carboxyl groups, carbamoyl groups,sulfo groups, amino groups, cyano groups, mono-lower alkylamino group,di-lower alkylamino group, hydroxyl groups and halogen atoms, or a groupof the formula --Y--R³, wherein Y represents a carbonyl group,thiocarbonyl group or sulfonyl group, and R³ represents a hydrogen atom,lower alkyl group, cycloalkyl group, cycloalkylalkyl group, aryl group,aralkyl group, lower alkoxy group, hydrazino group, amino group,arylamino group, carbamoyl group or heterocyclic group, wherein thelower alkyl group, cycloalkyl group, cycloalkylalkyl group, aryl group,aralkyl group and heterocyclic group may each have 1 to 4 substituentsselected from the group consisting of halogen atoms, optionallyprotected hydroxyl groups, amino groups, carboxyl groups, carbamoylgroups, cyano groups and lower alkoxycarbonyl groups, and the aminogroup and carbamoyl group may each be mono- or di-substituted by loweralkyl group(s) optionally substituted by group(s) selected from thegroup consisting of halogen atoms, hydroxyl groups, amino groups,carboxyl groups, carbamoyl groups and lower alkoxycarbonyl groups; orR¹³ and R²³ combine together with the nitrogen atom to which they bindto form a heterocyclic group, wherein the heterocyclic group may have onthe ring lower alkyl group(s) which may be substituted by groupsselected from the group consisting of amino groups, hydroxyl groups,carboxyl groups and sulfo groups, wherein when R¹³ and/or R²³ contain afunctional group, each functional group is protected, optionally,removing protective group(s) existing in the product, to prepare acompound represented by formula (Ic) ##STR82## wherein R¹³, R²³, X¹, X²and G have the same meanings as defined above;or either formylating,alkylating, alkenylating, alkynylating, aralkylating, carbamoylating,thiocarbamoylating, alkanoylating or sulfonylating the amino group##STR83## of the compound of the above formula (Ic) wherein anyfunctional groups may be protected when R¹³ and R²³ represent a hydrogenatom, or condensing the above compound with a compound represented bythe following formula (V) or a derivative thereof wherein a functionalgroup is protected

    OHC--R.sup.6                                               (V)

wherein R⁶ represents a hydrogen atom or carboxyl group, or a loweralkyl group which may have 1 to 4 substituents selected from the groupconsisting of amino groups, mono-lower alkylamino groups, di-loweralkylamino group, hydroxyl groups, carboxyl groups and sulfo groups,and,if necessary, removing the protective groups existing in the product; orreducing the double bonds of the compound of the above formula (Ic) whenR¹³ and/or R²³ contain a double bond, or the compound prepared bycondensing the compound of the formula (Ic) with the compound of theformula (V) or the derivative thereof wherein the functional groups areprotected, and if necessary removing the protective group(s) existing inthe product; and if necessary, converting the resulting compound of theformula (I) into a pharmaceutically acceptable salt.
 2. The process ofclaim 1 wherein the compound of formula (II) is used.
 3. The process ofclaim 1 wherein the compound of formula (III) is used.